Hydrazide-containing hepatitis c serine protease inhibitors

ABSTRACT

The present invention relates to novel hydrazide-containing compounds of Formula I or Formula II, including pharmaceutically acceptable salts, esters, or prodrugs thereof which inhibit serine protease activity, particularly the activity of hepatitis C virus (HCV) NS3-NS4A protease. Consequently, the compounds of the present invention interfere with the life cycle of the hepatitis C virus and are also useful as antiviral agents. The present invention further relates to pharmaceutical compositions comprising the aforementioned compounds for administration to a subject suffering from HCV infection. The invention also relates to methods of treating an HCV infection in a subject by administering a pharmaceutical composition comprising the compounds of the present invention.

TECHNICAL FIELD

The present invention relates to novel hepatitis C virus (HCV) proteaseinhibitor compounds having antiviral activity against HCV, which arealso useful in the treatment of HCV infections. More specifically, theinvention relates to novel, hydrazide-containing HCV protease inhibitorcompounds, compositions containing such compounds and methods for usingthe same, as well as processes for making such compounds.

BACKGROUND OF THE INVENTION

HCV is the principal cause of non-A, non-B hepatitis and is anincreasingly severe public health problem both in the developed anddeveloping world. It is estimated that the virus infects over 200million people worldwide, surpassing the number of individuals infectedwith the human immunodeficiency virus (HIV) by nearly five-fold. HCVinfected patients, due to the high percentage of individuals inflictedwith chronic infections, are at an elevated risk of developing cirrhosisof the liver, subsequent hepatocellular carcinoma and terminal liverdisease. HCV is the most prevalent cause of hepatocellular cancer andcause of patients requiring liver transplantations in the western world.

There are considerable barriers to the development of anti-HCVtherapeutics, which include, but are not limited to, the persistence ofthe virus, the genetic diversity of the virus during replication in thehost, the high incident rate of the virus developing drug-resistantmutants, and the lack of reproducible infectious culture systems andsmall-animal models for HCV replication and pathogenesis. In a majorityof cases, given the mild course of the infection and the complex biologyof the liver, careful consideration must be given to antiviral drugs,which are likely to have significant side effects.

Only two approved therapies for HCV infection are currently available.The original treatment regimen generally involves a 3-12 month course ofintravenous interferon-α (IFN-α), while a new approved second-generationtreatment involves co-treatment with IFN-α and general antiviralnucleoside mimics like ribavirin. Both of these treatments suffer frominterferon related side effects as well as low efficacy against HCVinfections. There exists a need for the development of effectiveantiviral agents for treatment of HCV infection due to the poortolerability and disappointing efficacy of existing therapies.

In a patient population where the majority of individuals arechronically infected and asymptomatic, and the prognoses are unknown, aneffective drug would desirably possess significantly fewer side effectsthan the currently available treatments. The hepatitis C non-structuralprotein-3 (NS3) is a proteolytic enzyme required for processing of theviral polyprotein and consequently viral replication. Despite the hugenumber of viral variants associated with HCV infection, the active siteof the NS3 protease remains highly conserved, thus making its inhibitionan attractive mode of intervention. Recent success in the treatment ofHIV with protease inhibitors supports the concept that the inhibition ofNS3 is a key target in the battle against HCV.

HCV is a flaviridae type RNA virus. The HCV genome is enveloped andcontains a single strand RNA molecule composed of circa 9600 base pairs.It encodes a polypeptide comprised of approximately 3010 amino acids.

The HCV polyprotein is processed by viral and host peptidase into 10discreet peptides, which serve a variety of functions. There are threestructural proteins: C, E1 and E2. The P7 protein is of unknown functionand is comprised of a highly variable sequence. There are sixnon-structural proteins. NS2 is a zinc-dependent metalloproteinase thatfunctions in conjunction with a portion of the NS3 protein. NS3incorporates two catalytic functions (separate from its association withNS2): a serine protease at the N-terminal end, which requires NS4A as acofactor, and an ATP-ase-dependent helicase function at the carboxylterminus. NS4A is a tightly associated but non-covalent cofactor of theserine protease.

The NS3-NS4A protease is responsible for cleaving four sites on theviral polyprotein. The NS3-NS4A cleavage is autocatalytic, occurring incis. The remaining three hydrolyses, NS4A-NS4B, NS4B-NS5A and NS5A-NS5Ball occur in trans. NS3 is a serine protease, which is structurallyclassified as a chymotrypsin-like protease. While the NS3 serineprotease possesses proteolytic activity by itself, the HCV proteaseenzyme is not an efficient enzyme in terms of catalyzing polyproteincleavage. It has been shown that a central hydrophobic region of theNS4A protein is required for this enhancement. The complex formation ofthe NS3 protein with NS4A seems necessary to the processing events,enhancing the proteolytic efficacy at all of the sites.

A general strategy for the development of antiviral agents is toinactivate virally encoded enzymes, including NS3, that are essentialfor the replication of the virus. Current efforts directed toward thediscovery of NS3 protease inhibitors were reviewed by S. Tan, A. Pause,Y. Shi, N. Sonenberg, Hepatitis C Therapeutics: Current Status andEmerging Strategies, Nature Rev. Drug Discov., 1, 867-881 (2002). Otherpatent disclosures describing the synthesis of HCV protease inhibitorsare: WO 00/59929 (2000); WO 99/07733 (1999); WO 00/09543 (2000); WO99/50230 (1999); U.S. Pat. No. 5,861,297 (1999); U.S. Pat. No. 0,037,998(2002); WO 03/099274 (2003); U.S. Pat. No. 0,267,018 (2005); WO06/043145 (2006); and WO 06/086381 (2006); U.S. Pat. No. 0,267,018(2005); WO 06/043145 (2006); WO 06/086381 (2006); WO 07/025,307 (2007);WO 06/020276 (2006); WO 07/015,824 (2007); WO 07/016,441 (2007); WO07/015,855 (2007); WO 07/015,787 (2007); WO 07/014,927 (2007); WO07/014,926 (2007); WO 07/014,925 (2007); WO 07/014,924 (2007); WO07/014,923 (2007); WO 07/014,922 (2007); WO 07/014,921 (2007); WO07/014,920 (2007); WO 07/014,919 (2007); WO 07/014,918 (2007); WO07/009,227 (2007); WO 07/008,657 (2007); WO 07/001,406 (2007); WO07/011,658 (2007); WO 07/009,109 (2007); WO 06/119061 (2006).

SUMMARY OF THE INVENTION

The present invention relates to novel hydrazide-containing compoundsincluding pharmaceutically acceptable salts, esters, or prodrugs thereofwhich inhibit serine protease activity, particularly the activity ofhepatitis C virus (HCV) NS3-NS4A protease. Consequently, the compoundsof the present invention interfere with the life cycle of the hepatitisC virus and are also useful as antiviral agents. The present inventionfurther relates to pharmaceutical compositions comprising theaforementioned compounds for administration to a subject suffering fromHCV infection. The invention also relates to methods of treating an HCVinfection in a subject by administering a pharmaceutical compositioncomprising the compounds of the present invention.

In one embodiment of the present invention there are disclosed compoundsrepresented by Formula I or Formula II, or pharmaceutically acceptablesalts, esters, or prodrugs thereof:

-   -   Wherein    -   A is selected from the following groups:        -   (i) —(C═O)—O—R¹;        -   (ii) —(C═O)—R¹;        -   (iii) —C(═O)—NH—R²;        -   (iv) —S(O)₂—R¹;        -   (v) —S(O)₂NHR²;    -    Wherein, R¹ is selected from the following groups:        -   (i) aryl;        -   (ii) substituted aryl;        -   (iii) heteroaryl;        -   (iv) substituted heteroaryl;        -   (v) heterocycloalkyl;        -   (vi) substituted heterocycloalkyl;        -   (vii) —C₁-C₈ alkyl, optionally containing 0, 1, 2, or 3            heteroatoms selected from O, S, or N;        -   (viii) —C₂-C₈ alkenyl, optionally containing 0, 1, 2, or 3            heteroatoms selected from O, S, or N;        -   (ix) —C₂-C₈ alkynyl, optionally containing 0, 1, 2, or 3            heteroatoms selected from O, S, or N;        -   (x) substituted —C₁-C₈ alkyl, optionally containing 0, 1, 2,            or 3 heteroatoms selected from O, S, or N;        -   (xi) substituted —C₂-C₈ alkenyl, optionally containing 0, 1,            2, or 3 heteroatoms selected from O, S, or N;        -   (xii) substituted —C₂-C₈ alkynyl, optionally containing 0,            1, 2, or 3 heteroatoms selected from O, S or N;        -   (xiii) —C₃-C₁₂ cycloalkyl;        -   (xiv) substituted —C₃-C₁₂ cycloalkyl;        -   (xv) —C₃-C₁₂ cycloalkenyl; and        -   (xvi) substituted —C₃-C₁₂ cycloalkenyl;    -    Wherein, R² is selected from the following groups:        -   (i) hydrogen;        -   (ii) —C₁-C₈ alkyl, optionally containing 0, 1, 2, or 3            heteroatoms selected from O, S, or N;        -   (iii) —C₂-C₈ alkenyl, optionally containing 0, 1, 2, or 3            heteroatoms selected from O, S, or N;        -   (iv) —C₂-C₈ alkynyl, optionally containing 0, 1, 2, or 3            heteroatoms selected from O, S, or N;        -   (v) substituted —C₁-C₈ alkyl, optionally containing 0, 1, 2,            or 3 heteroatoms selected from O, S, or N;        -   (vi) substituted —C₂-C₈ alkenyl, optionally containing 0, 1,            2, or 3 heteroatoms selected from O, S, or N;        -   (vii) substituted —C₂-C₈ alkynyl, optionally containing 0,            1, 2, or 3 heteroatoms selected from O, S or N;    -   L is selected from the following groups:        -   (i) —C₁-C₈ alkyl, optionally containing 0, 1, 2, or 3            heteroatoms selected from O, S, or N;        -   (ii) —C₂-C₈ alkenyl, optionally containing 0, 1, 2, or 3            heteroatoms selected from O, S, or N;        -   (iii) —C₂-C₈ alkynyl, optionally containing 0, 1, 2, or 3            heteroatoms selected from O, S or N;        -   (iv) substituted —C₁-C₈ alkyl, optionally containing 0, 1,            2, or 3 heteroatoms selected from O, S, or N;        -   (v) substituted —C₂-C₈ alkenyl, optionally containing 0, 1,            2, or 3 heteroatoms selected from O, S, or N;        -   (vi) substituted —C₂-C₈ alkynyl, optionally containing 0, 1,            2, or 3 heteroatoms selected from O, S or N;    -   =a bond connected to an undefined stereogenic center;    -   X is absent or is selected from the following groups:        -   (i) O;        -   (ii) S;        -   (iii) NR³;    -    Wherein, R³ is selected from the following groups:        -   (i) hydrogen;        -   (ii) —C₁-C₈ alkyl, optionally containing 0, 1, 2, or 3            heteroatoms selected from O, S, or N;        -   (iii) —C₃-C₁₂ cycloalkyl;        -   (iv) substituted —C₃-C₁₂ cycloalkyl;    -   Y is absent or is selected from the following groups:        -   (i) —C₁-C₆ alkyl, optionally containing 0, 1, 2, or 3            heteroatoms selected from O, S, or N, optionally substituted            with one or more substituent selected from halogen, aryl,            substituted aryl, heteroaryl, or substituted heteroaryl;        -   (ii) —C₂-C₆ alkenyl, optionally containing 0, 1, 2, or 3            heteroatoms selected from O, S, or N, optionally substituted            with one or more substituent selected from halogen, aryl,            substituted aryl, heteroaryl, or substituted heteroaryl;        -   (iii) —C₂-C₆ alkynyl, optionally containing 0, 1, 2, or 3            heteroatoms selected from O, S, or N, optionally substituted            with one or more substituent selected from halogen, aryl,            substituted aryl, heteroaryl, or substituted heteroaryl; and        -   (iv) —C₃-C₁₂ cycloalkyl, optionally substituted —C₃-C₁₂            cycloalkyl, heterocycloalkyl, substituted heterocycloalkyl;        -   (v) —C═O;        -   (vi) —C═S;        -   (vii) —SO;        -   (viii) —SO₂;    -   Z is selected from the following groups:        -   (i) aryl or alkylaryl optionally substituted with R⁴;        -   (ii) heteroaryl or alkylheteroaryl optionally substituted            with R⁴;        -   (iii) —N═CR⁵R⁶;    -    Wherein, R⁴ is selected from the following groups:        -   (i) C₁₋₆ alkyl;        -   (ii) C₃₋₇ cycloalkyl;        -   (iii) C₁₋₆ alkoxy;        -   (iv) C₃₋₇ cycloalkoxy;        -   (v) halo-C₁₋₆ alkyl;        -   (vi) CF₃;        -   (vii) mono- or di-halo-C₁₋₆ alkoxy;        -   (viii) cyano;        -   (ix) halo;        -   (x) thioalkyl;        -   (xi) hydroxyl;        -   (xii) alkanoyl;        -   (xiii) NO₂;        -   (xiv) SH;        -   (xv) amino;        -   (xvi) C₁₋₆ alkylamino;        -   (xvii) di(C₁₋₆) alkylamino;        -   (xviii) di(C₁₋₆)alkylamide;        -   (xix) carboxyl;        -   (xx) (C₁₋₆) carboxyester;        -   (xxi) (C₁₋₆) alkylsulfone;        -   (xxii) (C₁₋₆) alkylsulfonamide;        -   (xxiii) di(C₁₋₆) alkyl(alkoxy)amine;        -   (xxiv) C₆₋₁₀ aryl;        -   (xxv) C₇₋₁₄ alkylaryl; or a        -   (xxvi) 5-7 membered monocyclic heterocycle;    -    Wherein, R⁵ and R⁶ are independently selected from the        following groups:        -   (i) aryl;        -   (ii) substituted aryl;        -   (iii) heteroaryl;        -   (iv) substituted heteroaryl; (taken together with the            nitrogen that they are attached to, R⁵ and R⁶ may also form            a ring);    -   m=0, 1, or 2;    -   n=1, 2, or 3;    -   Q is selected from the following groups:        -   (i) hydrogen;        -   (ii) SR⁷;        -   (iii) —C₁-C₈ alkyl, optionally containing 0, 1, 2, or 3            heteroatoms selected from O, S, or N;        -   (iv) —C₂-C₈ alkenyl, optionally containing 0, 1, 2, or 3            heteroatoms selected from O, S, or N;        -   (v) —C₂-C₈ alkynyl, optionally containing 0, 1, 2, or 3            heteroatoms selected from O, S or N;        -   (vi) substituted —C₁-C₈ alkyl, optionally containing 0, 1,            2, or 3 heteroatoms selected from O, S, or N;        -   (vii) substituted —C₂-C₈ alkenyl, optionally containing 0,            1, 2, or 3 heteroatoms selected from O, S, or N;        -   (viii) substituted —C₂-C₈ alkynyl containing 0, 1, 2, or 3            heteroatoms selected from O, S or N;        -   (ix) —C₃-C₁₂ cycloalkyl;        -   (x) substituted —C₃-C₁₂ cycloalkyl;        -   (xi) —C₃-C₁₂ cycloalkenyl;        -   (xii) substituted —C₃-C₁₂ cycloalkenyl;        -   (xiii) heterocyclic; and        -   (xiv) substituted heterocyclic;    -    Wherein, R⁷ is selected from the following groups:        -   (i) aryl;        -   (ii) substituted aryl;        -   (iii) heteroaryl;        -   (iv) substituted heteroaryl;        -   (v) heterocycloalkyl;        -   (vi) substituted heterocycloalkyl;        -   (vii) —C₁-C₈ alkyl, optionally containing 0, 1, 2, or 3            heteroatoms selected from O, S, or N;        -   (viii) —C₂-C₈ alkenyl, optionally containing 0, 1, 2, or 3            heteroatoms selected from O, S, or N;        -   (ix) —C₂-C₈ alkynyl, optionally containing 0, 1, 2, or 3            heteroatoms selected from O, S, or N;        -   (x) substituted —C₁-C₈ alkyl, optionally containing 0, 1, 2,            or 3 heteroatoms selected from O, S, or N;        -   (xi) substituted —C₂-C₈ alkenyl, optionally containing 0, 1,            2, or 3 heteroatoms selected from O, S, or N;        -   (xii) substituted —C₂-C₈ alkynyl, optionally containing 0,            1, 2, or 3 heteroatoms selected from O, S or N;        -   (xiii) —C₃-C₁₂ cycloalkyl;        -   (xiv) substituted —C₃-C₁₂ cycloalkyl;        -   (xv) —C₃-C₁₂ cycloalkenyl; and        -   (xvi) substituted —C₃-C₁₂ cycloalkenyl;    -   s=0, 1, 2, or 3;    -   V and U are independently selected from the following groups:        -   (i) hydrogen;        -   (ii) —(C═O)—O—R⁸;        -   (iii) —(C═O)—R⁹;        -   (iv) —C(═O)—NR¹⁰R¹¹; (taken together with the nitrogen that            they are attached to, U and V may also form a ring);    -    Wherein, R⁸ and R⁹ are selected from the following groups:        -   (i) aryl;        -   (ii) substituted aryl;        -   (iii) heteroaryl;        -   (iv) substituted heteroaryl;        -   (v) heterocycloalkyl;        -   (vi) substituted heterocycloalkyl;        -   (vii) —C₁-C₈ alkyl, optionally containing 0, 1, 2, or 3            heteroatoms selected from O, S, or N;        -   (viii) —C₂-C₈ alkenyl, optionally containing 0, 1, 2, or 3            heteroatoms selected from O, S, or N;        -   (ix) —C₂-C₈ alkynyl, optionally containing 0, 1, 2, or 3            heteroatoms selected from O, S, or N;        -   (x) substituted —C₁-C₈ alkyl, optionally containing 0, 1, 2,            or 3 heteroatoms selected from O, S, or N;        -   (xi) substituted —C₂-C₈ alkenyl, optionally containing 0, 1,            2, or 3 heteroatoms selected from O, S, or N;        -   (xii) substituted —C₂-C₈ alkynyl, optionally containing 0,            1, 2, or 3 heteroatoms selected from O, S or N;        -   (xiii) —C₃-C₁₂ cycloalkyl;        -   (xiv) substituted —C₃-C₁₂ cycloalkyl;        -   (xv) —C₃-C₁₂ cycloalkenyl; and        -   (xvi) substituted —C₃-C₁₂ cycloalkenyl;    -    Wherein, R¹⁰ and R¹¹ are selected from the following groups:        -   (i) hydrogen;        -   (ii) aryl;        -   (iii) substituted aryl;        -   (iv) heteroaryl;        -   (v) substituted heteroaryl;        -   (vi) heterocycloalkyl;        -   (vii) substituted heterocycloalkyl;        -   (viii) —C₁-C₈ alkyl, optionally containing 0, 1, 2, or 3            heteroatoms selected from O, S, or N;        -   (ix) —C₂-C₈ alkenyl, optionally containing 0, 1, 2, or 3            heteroatoms selected from O, S, or N;        -   (x) —C₂-C₈ alkynyl, optionally containing 0, 1, 2, or 3            heteroatoms selected from O, S, or N;        -   (xi) substituted —C₁-C₈ alkyl, optionally containing 0, 1,            2, or 3 heteroatoms selected from O, S, or N;        -   (xii) substituted —C₂-C₈ alkenyl, optionally containing 0,            1, 2, or 3 heteroatoms selected from O, S, or N;        -   (xiii) substituted —C₂-C₈ alkynyl, optionally containing 0,            1, 2, or 3 heteroatoms selected from O, S or N;        -   (xiv) —C₃-C₁₂ cycloalkyl;        -   (xv) substituted —C₃-C₁₂ cycloalkyl;        -   (xvi) —C₃-C₁₂ cycloalkenyl; and        -   (xvii) substituted —C₃-C₁₂ cycloalkenyl; (taken together            with the nitrogen that they are attached to, R¹⁰ and R¹¹ may            also form a ring);    -   W is selected from —CH₂—, —O—, —S—, —S(O)₂—, —CO—, —C(O)O—,        —C(O)NH—, —CHF—, —CF₂—, aryl, substituted aryl, heteroaryl, and        substituted heteroaryl;    -   =either a carbon-carbon single bond or a carbon-carbon double        bond    -   j=0, 1, 2, 3, or 4;    -   k=1, 2, or 3.

DETAILED DESCRIPTION OF THE INVENTION

A first embodiment of the invention is a compound represented by eitherFormula I or Formula II, as described above, or a pharmaceuticallyacceptable salt, ester or prodrug thereof, alone or in combination witha pharmaceutically acceptable carrier or excipient.

Representative subgenera of the invention include, but are not limitedto compounds of Formula III or Formula IV:

-   -   Wherein A, L, j, k, and V are all as previously defined;    -   J is either absent or ═O, S, NR¹², CO, (CO)NR¹², (CO)O,        NR¹²(CO), NH(CO)NH, NR¹²SO₂;    -    Wherein, R¹² is selected from the following groups:        -   (i) hydrogen;        -   (ii) —C₁-C₈ alkyl, optionally containing 0, 1, 2, or 3            heteroatoms selected from O, S, or N;        -   (iii) —C₃-C₁₂ cycloalkyl;        -   (iv) substituted —C₃-C₁₂ cycloalkyl;    -   R¹³ is selected from aryl, substituted aryl, heteroaryl, and        substituted heteroaryl;    -   R⁸¹, R⁸², R⁸³, and R⁸⁴ are independently chosen from the        following groups:        -   (i) hydrogen;        -   (ii) halogen;        -   (iii) —NO₂;        -   (iv) —CN;        -   (v) MR¹⁴, wherein M is absent, or ═O, S, NR¹⁰R¹¹, wherein            R¹⁰ and R¹¹ are as previously defined;        -   (vi) aryl;        -   (vii) substituted aryl;        -   (viii) heteroaryl;        -   (ix) substituted heteroaryl;        -   (x) heterocycloalkyl; and        -   (xi) substituted heterocycloalkyl;    -    Wherein, R¹⁴ is selected from the following groups:        -   (i) hydrogen;        -   (ii) aryl;        -   (iii) substituted aryl;        -   (iv) heteroaryl;        -   (v) substituted heteroaryl;        -   (vi) heterocycloalkyl;        -   (vii) substituted heterocycloalkyl;        -   (viii) —C₁-C₈ alkyl, optionally containing 0, 1, 2, or 3            heteroatoms selected from O, S, or N;        -   (ix) —C₂-C₈ alkenyl, optionally containing 0, 1, 2, or 3            heteroatoms selected from O, S, or N;        -   (x) —C₂-C₈ alkynyl, optionally containing 0, 1, 2, or 3            heteroatoms selected from O, S, or N;        -   (xi) substituted —C₁-C₈ alkyl, optionally containing 0, 1,            2, or 3 heteroatoms selected from O, S, or N;        -   (xii) substituted —C₂-C₈ alkenyl, optionally containing 0,            1, 2, or 3 heteroatoms selected from O, S, or N;        -   (xiii) substituted —C₂-C₈ alkynyl, optionally containing 0,            1, 2, or 3 heteroatoms selected from O, S or N;

Representative subgenera of the invention also include, but are notlimited to compounds of Formula V or Formula VI:

-   -   Wherein A, L, j, k, V, R⁸¹, R³², R⁸³, and R⁸⁴ are all as        previously defined;        Representative subgenera of the invention also include, but are        not limited to compounds of Formula VII or Formula VIII:

-   -   wherein A, L, j, k, V, J, R¹³, R⁸¹, R⁸², R⁸³, and R⁸⁴ are all as        previously defined;    -   Representative compounds of the invention include, but are not        limited to, compounds 1-113 of Formula IX:

-   -   Wherein A, P*(X—Y-Z), and V are delineated for each example as        set forth in Table 1:

TABLE 1 EXAMPLE # A P* V 1

2

3

H 4

H 5

6

7

8

9

10

11

12

13

14

15

16

17

18

19

20

21

22

23

24

25

26

27

28

29

30

31

32

33

34

35

36

37

38

39

40

41

42

43

44

45

46

47

48

49

50

51

52

53

54

55

56

57

58

59

60

61

62

63

64

65

66

67

68

69

70

71

72

73

74

75

76

77

78

79

80

81

82

83

84

85

86

87

88

89

90

91

92

93

94

95

96

97

98

99

100

101

102

103

104

105

106

107

108

109

110

111

112

113

Representative compounds of the invention also include, but are notlimited to, compounds 114-226 of Formula X:

Wherein A, P*(X—Y-Z), V, and L are delineated for each example as setforth in Table 2:

TABLE 2 EXAMPLE # A P* V L 114

115

116

117

118

119

120

121

122

123

124

125

126

127

128

129

130

131

132

133

134

135

136

137

138

139

140

141

142

143

144

145

146

147

148

149

150

151

152

153

154

155

156

157

158

159

160

161

162

163

164

165

166

167

168

169

170

171

172

173

174

175

176

177

178

179

180

181

182

183

184

185

186

187

188

189

190

191

192

193

194

195

196

197

198

199

200

201

202

203

204

205

206

207

208

209

210

211

212

213

214

215

216

217

218

219

220

221

222

223

224

225

226

A further embodiment of the present invention includes pharmaceuticalcompositions comprising any single compound delineated herein, or apharmaceutically acceptable salt, ester, or prodrug thereof, with apharmaceutically acceptable carrier or excipient.

Yet another embodiment of the present invention is a pharmaceuticalcomposition comprising a combination of two or more compounds delineatedherein, or a pharmaceutically acceptable salt, ester, or prodrugthereof, with a pharmaceutically acceptable carrier or excipient.

According to an alternate embodiment, the pharmaceutical compositions ofthe present invention may further contain other anti-HCV agents.Examples of anti-HCV agents include, but are not limited to,α-interferon, β-interferon, ribavirin, and amantadine. For furtherdetails see S. Tan, A. Pause, Y. Shi, N. Sonenberg, Hepatitis CTherapeutics: Current Status and Emerging Strategies, Nature Rev. DrugDiscov., 1, 867-881 (2002); WO 00/59929 (2000); WO 99/07733 (1999); WO00/09543 (2000); WO 99/50230 (1999); U.S. Pat. No. 5,861,297 (1999); andUS2002/0037998 (2002) which are herein incorporated by reference intheir entirety.

According to an additional embodiment, the pharmaceutical compositionsof the present invention may further contain other HCV proteaseinhibitors.

According to yet another embodiment, the pharmaceutical compositions ofthe present invention may further comprise inhibitor(s) of other targetsin the HCV life cycle, including, but not limited to, helicase,polymerase, metalloprotease, and internal ribosome entry site (IRES).

According to another embodiment, the present invention includes methodsof treating hepatitis C infections in a subject in need of suchtreatment by administering to said subject a therapeutically effectiveamount of the pharmaceutical compounds or compositions of the presentinvention. The methods can further include administration of anadditional therapeutic agent, including another antiviral agent or ananti-HCV agent. The additional agent can be co-administered,concurrently administered or sequentially administered with the compoundor composition delineated herein. The methods herein can further includethe step of identifying that the subject is in need of treatment forhepatitis C infection. The identification can be by subjective (e.g.,health care provider determination) or objective (e.g., diagnostic test)means.

An additional embodiment of the present invention includes methods oftreating biological samples by contacting the biological samples withthe compounds of the present invention.

Yet a further aspect of the present invention is a process of making anyof the compounds delineated herein employing any of the synthetic meansdelineated herein.

Definitions

Listed below are definitions of various terms used to describe thisinvention. These definitions apply to the terms as they are usedthroughout this specification and claims, unless otherwise limited inspecific instances, either individually or as part of a larger group.

The terms “C₁-C₆ alkyl,” or “C₁-C₈ alkyl,” as used herein, refer tosaturated, straight- or branched-chain hydrocarbon radicals containingbetween one and six, or one and eight carbon atoms, respectively.Examples of C₁-C₆ alkyl radicals include, but are not limited to,methyl, ethyl, propyl, isopropyl, n-butyl, tert-butyl, neopentyl,n-hexyl radicals; and examples of C₁-C₈ alkyl radicals include, but arenot limited to, methyl, ethyl, propyl, isopropyl, n-butyl, tert-butyl,neopentyl, n-hexyl, heptyl, octyl radicals.

The terms “C₂-C₆ alkenyl,” or “C₂-C₈ alkenyl,” as used herein, denote amonovalent group derived from a hydrocarbon moiety containing from twoto six, or two to eight carbon atoms having at least one carbon-carbondouble bond by the removal of a single hydrogen atom. Alkenyl groupsinclude, but are not limited to, for example, ethenyl, propenyl,butenyl, 1-methyl-2-buten-1-yl, heptenyl, octenyl and the like.

The term “C₂-C₆ alkynyl,” or “C₂-C₈ alkynyl,” as used herein, denote amonovalent group derived from a hydrocarbon moiety containing from twoto six, or two to eight carbon atoms having at least one carbon-carbontriple bond by the removal of a single hydrogen atom. Representativealkynyl groups include, but are not limited to, for example, ethynyl,1-propynyl, 1-butynyl, heptynyl, octynyl and the like.

The term “C₃-C₈-cycloalkyl”, or “C₃-C₁₂-cycloalkyl,” as used herein,denotes a monovalent group derived from a monocyclic or polycyclicsaturated carbocyclic ring compound by the removal of a single hydrogenatom, respectively. Examples of C₃-C₈-cycloalkyl include, but notlimited to, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl,cyclopentyl and cyclooctyl; and examples of C₃-C₁₂-cycloalkyl include,but not limited to, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl,bicyclo[2.2.1]heptyl, and bicyclo[2.2.2]octyl.

The term “C₃-C₈-cycloalkenyl”, or “C₃-C₁₂-cycloalkenyl” as used herein,denote a monovalent group derived from a monocyclic or polycycliccarbocyclic ring compound having at least one carbon-carbon double bondby the removal of a single hydrogen atom. Examples of C₃-C₈-cycloalkenylinclude, but not limited to, cyclopropenyl, cyclobutenyl, cyclopentenyl,cyclohexenyl, cycloheptenyl, cyclooctenyl, and the like; and examples ofC₃-C₁₂-cycloalkenyl include, but not limited to, cyclopropenyl,cyclobutenyl, cyclopentenyl, cyclohexenyl, cycloheptenyl, cyclooctenyl,and the like.

The term “aryl,” as used herein, refers to a mono- or polycycliccarbocyclic ring system having one or more aromatic rings including, butnot limited to, phenyl, naphthyl, tetrahydronaphthyl, indanyl, idenyland the like.

The term “arylalkyl,” as used herein, refers to a C₁-C₃ alkyl or C₁-C₆alkyl residue attached to an aryl ring. Examples include, but are notlimited to, benzyl, phenethyl and the like.

The term “heteroaryl,” as used herein, refers to a mono- or polycyclic(e.g. bi-, or tri-cyclic or more), fused or non-fused, aromatic radicalor ring having from five to ten ring atoms of which one or more ringatom is selected from, for example, S, O and N; zero, one or two ringatoms are additional heteroatoms independently selected from, forexample, S, O and N; and the remaining ring atoms are carbon, whereinany N or S contained within the ring may be optionally oxidized.Heteroaryl includes, but is not limited to, pyridinyl, pyrazinyl,pyrimidinyl, pyrrolyl, pyrazolyl, imidazolyl, thiazolyl, oxazolyl,isooxazolyl, thiadiazolyl, oxadiazolyl, thiophenyl, furanyl, quinolinyl,isoquinolinyl, benzimidazolyl, benzooxazolyl, quinoxalinyl, and thelike.

The term “heteroarylalkyl,” as used herein, refers to a C₁-C₃ alkyl orC₁-C₆ alkyl residue attached to a heteroaryl ring. Examples include, butare not limited to, pyridinylmethyl, pyrimidinylethyl and the like.

The term “heterocycloalkyl,” as used herein, refers to a non-aromatic3-, 4-, 5-, 6- or 7-membered ring or a bi- or tri-cyclic group fusedsystem, where (i) each ring contains between one and three heteroatomsindependently selected from oxygen, sulfur and nitrogen, (ii) each5-membered ring has 0 to 1 double bonds and each 6-membered ring has 0to 2 double bonds, (iii) the nitrogen and sulfur heteroatoms mayoptionally be oxidized, (iv) the nitrogen heteroatom may optionally bequaternized, and (iv) any of the above rings may be fused to a benzenering. Representative heterocycloalkyl groups include, but are notlimited to, [1,3]dioxolane, pyrrolidinyl, pyrazolinyl, pyrazolidinyl,imidazolinyl, imidazolidinyl, piperidinyl, piperazinyl, oxazolidinyl,isoxazolidinyl, morpholinyl, thiazolidinyl, isothiazolidinyl, andtetrahydrofuryl.

The terms “substituted”, “substituted C₁-C₆ alkyl,” “substituted C₁-C₈alkyl,” “substituted C₂-C₆ alkenyl,” “substituted C₂-C₈ alkenyl,”“substituted C₂-C₆ alkynyl”, “substituted C₂-C₈ alkynyl”, “substitutedC₃-C₁₂ cycloalkyl,” “substituted C₃-C₈ cycloalkenyl,” “substitutedC₃-C₁₂ cycloalkenyl,” “substituted aryl”, “substituted heteroaryl,”“substituted arylalkyl”, “substituted heteroarylalkyl,” “substitutedheterocycloalkyl,” as used herein, refer to CH, NH, C₁-C₆ alkyl, C₁-C₈alkyl, C₂-C₆ alkenyl, C₂-C₈ alkenyl, C₂-C₆ alkynyl, C₂-C₈ alkynyl,C₃-C₁₂ cycloalkyl, C₃-C₈ cycloalkenyl, C₃-C₁₂ cycloalkenyl, aryl,heteroaryl, arylalkyl, heteroarylalkyl, heterocycloalkyl groups aspreviously defined, substituted by independent replacement of one, two,or three or more of the hydrogen atoms thereon with substituentsincluding, but not limited to, —F, —Cl, —Br, —I, —OH, protected hydroxy,—NO₂, —CN, —NH₂, protected amino, —NH-C₁-C₁₂-alkyl, —NH-C₂-C₁₂-alkenyl,—NH—C₂-C₁₂-alkenyl, —NH—C₃-C₁₂-cycloalkyl, —NH-aryl, —NH-heteroaryl,—NH-heterocycloalkyl, -dialkylamino, -diarylamino, -diheteroarylamino,—O—C₁-C₁₂-alkyl, —O—C₂-C₁₂-alkenyl, —O—C₂-C₁₂-alkenyl,—O—C₃-C₁₂-cycloalkyl, —O-aryl, —O-heteroaryl, —O-heterocycloalkyl,—C(O)—C₁-C₁₂-alkyl, —C(O)—C₂-C₁₂-alkenyl, —C(O)—C₂-C₁₂-alkenyl,—C(O)—C₃-C₁₂-cycloalkyl, —C(O)-aryl, —C(O)-heteroaryl,—C(O)-heterocycloalkyl, —CONH₂, —CONH—C₁-C₁₂-alkyl,—CONH—C₂-C₁₂-alkenyl, —CONH—C₂-C₁₂-alkenyl, —CONH—C₃-C₁₂-cycloalkyl,—CONH-aryl, —CONH-heteroaryl, —CONH-heterocycloalkyl,—OCO₂—C₁-C₁₂-alkyl, —OCO₂—C₂-C₁₂-alkenyl, —OCO₂—C₂-C₁₂-alkenyl,—OCO₂—C₃-C₁₂-cycloalkyl, —OCO₂-aryl, —OCO₂-heteroaryl,—OCO₂-heterocycloalkyl, —OCONH₂, —OCONH—C₁-C₁₂-alkyl,—OCONH—C₂-C₁₂-alkenyl, —OCONH—C₂-C₁₂-alkenyl, —OCONH—C₃-C₁₂-cycloalkyl,—OCONH-aryl, —OCONH-heteroaryl, —OCONH-heterocycloalkyl,—NHC(O)—C₁-C₁₂-alkyl, —NHC(O)—C₂-C₁₂-alkenyl, —NHC(O)—C₂-C₁₂-alkenyl,—NHC(O)—C₃-C₁₂-cycloalkyl, —NHC(O)-aryl, —NHC(O)-heteroaryl,—NHC(O)-heterocycloalkyl, —NHCO₂—C₁-C₁₂-alkyl, —NHCO₂—C₂-C₁₂-alkenyl,—NHCO₂—C₂-C₁₂-alkenyl, —NHCO₂—C₃-C₁₂-cycloalkyl, —NHCO₂-aryl,—NHCO₂-heteroaryl, —NHCO₂-heterocycloalkyl, —NHC(O)NH₂,—NHC(O)NH—C₁-C₁₂-alkyl, —NHC(O)NH—C₂-C₁₂-alkenyl,—NHC(O)NH—C₂-C₁₂-alkenyl, —NHC(O)NH—C₃-C₁₂-cycloalkyl, —NHC(O)NH-aryl,—NHC(O)NH-heteroaryl, —NHC(O)NH-heterocycloalkyl, NHC(S)NH₂,—NHC(S)NH—C₁-C₁₂-alkyl, —NHC(S)NH—C₂-C₁₂-alkenyl,—NHC(S)NH—C₂-C₁₂-alkenyl, —NHC(S)NH—C₃-C₁₂-cycloalkyl, —NHC(S)NH-aryl,—NHC(S)NH-heteroaryl, —NHC(S)NH-heterocycloalkyl, —NHC(NH)NH₂,—NHC(NH)NH—C₁-C₁₂-alkyl, —NHC(NH)NH—C₂-C₁₂-alkenyl,—NHC(NH)NH—C₂-C₁₂-alkenyl, —NHC(NH)NH—C₃-C₁₂-cycloalkyl,—NHC(NH)NH-aryl, —NHC(NH)NH-heteroaryl, —NHC(NH)NH-heterocycloalkyl,—NHC(NH)—C₁-C₁₂-alkyl, —NHC(NH)—C₂-C₁₂-alkenyl, —NHC(NH)—C₂-C₁₂-alkenyl,—NHC(NH)—C₃-C₁₂-cycloalkyl, —NHC(NH)-aryl, —NHC(NH)-heteroaryl,—NHC(NH)-heterocycloalkyl, —C(NH)NH—C₁-C₁₂-alkyl,—C(NH)NH—C₂-C₁₂-alkenyl, —C(NH)NH—C₂-C₁₂-alkenyl,—C(NH)NH—C₃-C₁₂-cycloalkyl, —C(NH)NH-aryl, —C(NH)NH-heteroaryl,—C(NH)NH-heterocycloalkyl, —S(O)—C₁-C₁₂-alkyl, —S(O)—C₂-C₁₂-alkenyl,—S(O)—C₂-C₁₂-alkenyl, —S(O)—C₃-C₁₂-cycloalkyl, —S(O)-aryl,—S(O)-heteroaryl, —S(O)-heterocycloalkyl-SO₂NH₂, —SO₂NH—C₁-C₁₂-alkyl,—SO₂NH—C₂-C₁₂-alkenyl, —SO₂NH—C₂-C₁₂-alkenyl, —SO₂NH—C₃-C₁₂-cycloalkyl,—SO₂NH-aryl, —SO₂NH-heteroaryl, —SO₂NH-heterocycloalkyl,—NHSO₂—C₁-C₁₂-alkyl, —NHSO₂—C₂-C₁₂-alkenyl, —NHSO₂—C₂-C₁₂-alkenyl,—NHSO₂—C₃-C₁₂-cycloalkyl, —NHSO₂-aryl, —NHSO₂-heteroaryl,—NHSO₂-heterocycloalkyl, —CH₂NH₂, —CH₂SO₂CH₃, -aryl, -arylalkyl,-heteroaryl, -heteroarylalkyl, -heterocycloalkyl, —C₃-C₁₂-cycloalkyl,polyalkoxyalkyl, polyalkoxy, -methoxymethoxy, -methoxyethoxy, —SH,—S—C₁-C₁₂-alkyl, —S—C₂-C₁₂-alkenyl, —S—C₂-C₁₂-alkenyl,—S—C₃-C₁₂-cycloalkyl, —S-aryl, —S-heteroaryl, —S-heterocycloalkyl, ormethylthiomethyl. It is understood that the aryls, heteroaryls, alkyls,and the like can be further substituted.

In accordance with the invention, any of the aryls, substituted aryls,heteroaryls and substituted heteroaryls described herein, can be anyaromatic group. Aromatic groups can be substituted or unsubstituted.

It is understood that any alkyl, alkenyl, alkynyl, cycloalkyl andcycloalkenyl moiety described herein can also be an aliphatic group, analicyclic group or a heterocyclic group. An “aliphatic group” isnon-aromatic moiety that may contain any combination of carbon atoms,hydrogen atoms, halogen atoms, oxygen, nitrogen or other atoms, andoptionally contain one or more units of unsaturation, e.g., doubleand/or triple bonds. An aliphatic group may be straight chained,branched or cyclic and preferably contains between about 1 and about 24carbon atoms, more typically between about 1 and about 12 carbon atoms.In addition to aliphatic hydrocarbon groups, aliphatic groups include,for example, polyalkoxyalkyls, such as polyalkylene glycols, polyamines,and polyimines, for example. Such aliphatic groups may be furthersubstituted. It is understood that aliphatic groups may be used in placeof the alkyl, alkenyl, alkynyl, alkylene, alkenylene, and alkynylenegroups described herein.

The term “alicyclic,” as used herein, denotes a monovalent group derivedfrom a monocyclic or polycyclic saturated carbocyclic ring compound bythe removal of a single hydrogen atom. Examples include, but not limitedto, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl,bicyclo[2.2.1]heptyl, and bicyclo[2.2.2]octyl. Such alicyclic groups maybe further substituted.

It will be apparent that in various embodiments of the invention, thesubstituted or unsubstituted alkyl, alkenyl, alkynyl, cycloalkyl,cycloalkenyl, cycloalkynyl, arylalkyl, heteroarylalkyl, andheterocycloalkyl are intended to be divalent or trivalent. Thus,alkylene, alkenylene, and alkynylene, cycloaklylene, cycloalkenylene,cycloalkynylene, arylalkylene, hetoerarylalkylene andheterocycloalkylene groups are to be included in the above definitions,and are applicable to provide the formulas herein with proper valency.

The terms “halo” and “halogen,” as used herein, refer to an atomselected from fluorine, chlorine, bromine and iodine.

The compounds described herein contain one or more asymmetric centersand thus give rise to enantiomers, diastereomers, and otherstereoisomeric forms that may be defined, in terms of absolutestereochemistry, as (R)- or (S)-, or as D- or L- for amino acids. Thepresent invention is meant to include all such possible isomers, as wellas their racemic and optically pure forms. Optical isomers may beprepared from their respective optically active precursors by theprocedures described above, or by resolving the racemic mixtures. Theresolution can be carried out in the presence of a resolving agent, bychromatography or by repeated crystallization or by some combination ofthese techniques, which are known to those skilled in the art. Furtherdetails regarding resolutions can be found in Jacques, et al.,Enantiomers, Racemates, and Resolutions (John Wiley & Sons, 1981). Whenthe compounds described herein contain olefinic double bonds or othercenters of geometric asymmetry, and unless specified otherwise, it isintended that the compounds include both E and Z geometric isomers.Likewise, all tautomeric forms are also intended to be included. Theconfiguration of any carbon-carbon double bond appearing herein isselected for convenience only and is not intended to designate aparticular configuration unless the text so states; thus a carbon-carbondouble bond depicted arbitrarily herein as trans may be cis, trans, or amixture of the two in any proportion.

The term “subject” as used herein refers to a mammal. A subjecttherefore refers to, for example, dogs, cats, horses, cows, pigs, guineapigs, and the like. Preferably the subject is a human. When the subjectis a human, the subject may be referred to herein as a patient.

As used herein, the term “pharmaceutically acceptable salt” refers tothose salts of the compounds formed by the process of the presentinvention which are, within the scope of sound medical judgment,suitable for use in contact with the tissues of humans and lower animalswithout undue toxicity, irritation, allergic response and the like, andare commensurate with a reasonable benefit/risk ratio. Pharmaceuticallyacceptable salts are well known in the art. For example, S. M. Berge, etal. describes pharmaceutically acceptable salts in detail in J.Pharmaceutical Sciences, 66: 1-19 (1977). The salts can be prepared insitu during the final isolation and purification of the compounds of theinvention, or separately by reacting the free base function with asuitable organic acid. Examples of pharmaceutically acceptable include,but are not limited to, nontoxic acid addition salts are salts of anamino group formed with inorganic acids such as hydrochloric acid,hydrobromic acid, phosphoric acid, sulfuric acid and perchloric acid orwith organic acids such as acetic acid, maleic acid, tartaric acid,citric acid, succinic acid or malonic acid or by using other methodsused in the art such as ion exchange. Other pharmaceutically acceptablesalts include, but are not limited to, adipate, alginate, ascorbate,aspartate, benzenesulfonate, benzoate, bisulfate, borate, butyrate,camphorate, camphorsulfonate, citrate, cyclopentanepropionate,digluconate, dodecylsulfate, ethanesulfonate, formate, fumarate,glucoheptonate, glycerophosphate, gluconate, hemisulfate, heptanoate,hexanoate, hydroiodide, 2-hydroxy-ethanesulfonate, lactobionate,lactate, laurate, lauryl sulfate, malate, maleate, malonate,methanesulfonate, 2-naphthalenesulfonate, nicotinate, nitrate, oleate,oxalate, palmitate, pamoate, pectinate, persulfate, 3-phenylpropionate,phosphate, picrate, pivalate, propionate, stearate, succinate, sulfate,tartrate, thiocyanate, p-toluenesulfonate, undecanoate, valerate salts,and the like. Representative alkali or alkaline earth metal saltsinclude sodium, lithium, potassium, calcium, magnesium, and the like.Further pharmaceutically acceptable salts include, when appropriate,nontoxic ammonium, quaternary ammonium, and amine cations formed usingcounterions such as halide, hydroxide, carboxylate, sulfate, phosphate,nitrate, alkyl having from 1 to 6 carbon atoms, sulfonate and arylsulfonate.

As used herein, the term “pharmaceutically acceptable ester” refers toesters of the compounds formed by the process of the present inventionwhich hydrolyze in vivo and include those that break down readily in thehuman body to leave the parent compound or a salt thereof. Suitableester groups include, for example, those derived from pharmaceuticallyacceptable aliphatic carboxylic acids, particularly alkanoic, alkenoic,cycloalkanoic and alkanedioic acids, in which each alkyl or alkenylmoiety advantageously has not more than 6 carbon atoms. Examples ofparticular esters include, but are not limited to, formates, acetates,propionates, butyrates, acrylates and ethylsuccinates.

The term “pharmaceutically acceptable prodrugs” as used herein refers tothose prodrugs of the compounds formed by the process of the presentinvention which are, within the scope of sound medical judgment,suitable for use in contact with the tissues of humans and lower animalswith undue toxicity, irritation, allergic response, and the like,commensurate with a reasonable benefit/risk ratio, and effective fortheir intended use, as well as the zwitterionic forms, where possible,of the compounds of the present invention. “Prodrug”, as used hereinmeans a compound, which is convertible in vivo by metabolic means (e.g.by hydrolysis) to afford any compound delineated by the formulae of theinstant invention. Various forms of prodrugs are known in the art, forexample, as discussed in Bundgaard, (ed.), Design of Prodrugs, Elsevier(1985); Widder, et al. (ed.), Methods in Enzymology, vol. 4, AcademicPress (1985); Krogsgaard-Larsen, et al., (ed). “Design and Applicationof Prodrugs, Textbook of Drug Design and Development, Chapter 5, 113-191(1991); Bundgaard, et al., Journal of Drug Deliver Reviews, 8:1-38(1992); Bundgaard, J. of Pharmaceutical Sciences, 77:285 et seq. (1988);Higuchi and Stella (eds.) Prodrugs as Novel Drug Delivery Systems,American Chemical Society (1975); and Bernard Testa & Joachim Mayer,“Hydrolysis In Drug And Prodrug Metabolism: Chemistry, Biochemistry AndEnzymology,” John Wiley and Sons, Ltd. (2002).

Combinations of substituents and variables envisioned by this inventionare only those that result in the formation of stable compounds. Theterm “stable”, as used herein, refers to compounds which possessstability sufficient to allow manufacture and which maintains theintegrity of the compound for a sufficient period of time to be usefulfor the purposes detailed herein (e.g., therapeutic or prophylacticadministration to a subject).

The synthesized compounds can be separated from a reaction mixture andfurther purified by a method such as column chromatography,high-performance liquid chromatography, or recrystallization. As usedherein, the term “substantially pure” for a compound refers to thephysical state of said compound after being obtained from a purificationprocess or processes described herein or that are well known to theskilled artisan, in sufficient purity to be characterizable by standardanalytical techniques described herein or as are well known to theskilled artisan.

In one embodiment, a substantially pure compound comprises a compound ofgreater than about 75% purity. This means that the compound does notcontain more than about 25% of any other compound. In one embodiment, asubstantially pure compound comprises a compound of greater than about80% purity. This means that the compound does not contain more thanabout 20% of any other compound. In one embodiment, a substantially purecompound comprises a compound of greater than about 85% purity. Thismeans that the compound does not contain more than about 15% of anyother compound. In one embodiment, a substantially pure compoundcomprises a compound of greater than about 90% purity. This means thatthe compound does not contain more than about 10% of any other compound.In another embodiment, a substantially pure compound comprises acompound of greater than about 95% purity. This means that the compounddoes not contain more than about 5% of any other compound. In anotherembodiment, a substantially pure compound comprises greater than about98% purity. This means that the compound does not contain more thanabout 2% of any other compound. In one embodiment, a substantially purecompound comprises a compound of greater than about 99% purity. Thismeans that the compound does not contain more than about 1% of any othercompound.

As can be appreciated by the skilled artisan, further methods ofsynthesizing the compounds of the formulae herein will be evident tothose of ordinary skill in the art. Additionally, the various syntheticsteps may be performed in an alternate sequence or order to give thedesired compounds. In addition, the solvents, temperatures, reactiondurations, etc. delineated herein are for purposes of illustration onlyand one of ordinary skill in the art will recognize that variation ofthe reaction conditions can produce the desired bridged macrocyclicproducts of the present invention. Synthetic chemistry transformationsand protecting group methodologies (protection and deprotection) usefulin synthesizing the compounds described herein are known in the art andinclude, for example, those such as described in R. Larock,Comprehensive Organic Transformations, VCH Publishers (1989); T. W.Greene and P. G. M. Wuts, Protective Groups in Organic Synthesis, 2d.Ed., John Wiley and Sons (1991); L. Fieser and M. Fieser, Fieser andFieser's Reagents for Organic Synthesis, John Wiley and Sons (1994); andL. Paquette, ed., Encyclopedia of Reagents for Organic Synthesis, JohnWiley and Sons (1995).

The compounds of this invention may be modified by appending variousfunctionalities via any synthetic means delineated herein to enhanceselective biological properties. Such modifications are known in the artand include those which increase biological penetration into a givenbiological system (e.g., blood, lymphatic system, central nervoussystem), increase oral availability, increase solubility to allowadministration by injection, alter metabolism and alter rate ofexcretion.

Pharmaceutical Compositions

The pharmaceutical compositions of the present invention comprise atherapeutically effective amount of a compound of the present inventionformulated together with one or more pharmaceutically acceptablecarriers. As used herein, the term “pharmaceutically acceptable carrier”means a non-toxic, inert solid, semi-solid or liquid filler, diluent,encapsulating material or formulation auxiliary of any type. Someexamples of materials which can serve as pharmaceutically acceptablecarriers are sugars such as lactose, glucose and sucrose; starches suchas corn starch and potato starch; cellulose and its derivatives such assodium carboxymethyl cellulose, ethyl cellulose and cellulose acetate;powdered tragacanth; malt; gelatin; talc; excipients such as cocoabutter and suppository waxes; oils such as peanut oil, cottonseed oil;safflower oil; sesame oil; olive oil; corn oil and soybean oil; glycols;such a propylene glycol; esters such as ethyl oleate and ethyl laurate;agar; buffering agents such as magnesium hydroxide and aluminumhydroxide; alginic acid; pyrogen-free water; isotonic saline; Ringer'ssolution; ethyl alcohol, and phosphate buffer solutions, as well asother non-toxic compatible lubricants such as sodium lauryl sulfate andmagnesium stearate, as well as coloring agents, releasing agents,coating agents, sweetening, flavoring and perfuming agents,preservatives and antioxidants can also be present in the composition,according to the judgment of the formulator. The pharmaceuticalcompositions of this invention can be administered to humans and otheranimals orally, rectally, parenterally, intracisternally,intravaginally, intraperitoneally, topically (as by powders, ointments,or drops), buccally, or as an oral or nasal spray.

Liquid dosage forms for oral administration include pharmaceuticallyacceptable emulsions, microemulsions, solutions, suspensions, syrups andelixirs. In addition to the active compounds, the liquid dosage formsmay contain inert diluents commonly used in the art such as, forexample, water or other solvents, solubilizing agents and emulsifierssuch as ethyl alcohol, isopropyl alcohol, ethyl carbonate, ethylacetate, benzyl alcohol, benzyl benzoate, propylene glycol, 1,3-butyleneglycol, dimethylformamide, oils (in particular, cottonseed, groundnut,corn, germ, olive, castor, and sesame oils), glycerol,tetrahydrofurfuryl alcohol, polyethylene glycols and fatty acid estersof sorbitan, and mixtures thereof. Besides inert diluents, the oralcompositions can also include adjuvants such as wetting agents,emulsifying and suspending agents, sweetening, flavoring, and perfumingagents.

Injectable preparations, for example, sterile injectable aqueous oroleaginous suspensions may be formulated according to the known artusing suitable dispersing or wetting agents and suspending agents. Thesterile injectable preparation may also be a sterile injectablesolution, suspension or emulsion in a nontoxic parenterally acceptablediluent or solvent, for example, as a solution in 1,3-butanediol. Amongthe acceptable vehicles and solvents that may be employed are water,Ringer's solution, U.S.P. and isotonic sodium chloride solution. Inaddition, sterile, fixed oils are conventionally employed as a solventor suspending medium. For this purpose any bland fixed oil can beemployed including synthetic mono- or diglycerides. In addition, fattyacids such as oleic acid are used in the preparation of injectables.

The injectable formulations can be sterilized, for example, byfiltration through a bacterial-retaining filter, or by incorporatingsterilizing agents in the form of sterile solid compositions which canbe dissolved or dispersed in sterile water or other sterile injectablemedium prior to use.

In order to prolong the effect of a drug, it is often desirable to slowthe absorption of the drug from subcutaneous or intramuscular injection.This may be accomplished by the use of a liquid suspension ofcrystalline or amorphous material with poor water solubility. The rateof absorption of the drug then depends upon its rate of dissolution,which, in turn, may depend upon crystal size and crystalline form.Alternatively, delayed absorption of a parenterally administered drugform is accomplished by dissolving or suspending the drug in an oilvehicle. Injectable depot forms are made by forming microencapsulematrices of the drug in biodegradable polymers such aspolylactide-polyglycolide. Depending upon the ratio of drug to polymerand the nature of the particular polymer employed, the rate of drugrelease can be controlled. Examples of other biodegradable polymersinclude poly(orthoesters) and poly(anhydrides). Depot injectableformulations are also prepared by entrapping the drug in liposomes ormicroemulsions which are compatible with body tissues.

Compositions for rectal or vaginal administration are preferablysuppositories which can be prepared by mixing the compounds of thisinvention with suitable non-irritating excipients or carriers such ascocoa butter, polyethylene glycol or a suppository wax which are solidat ambient temperature but liquid at body temperature and therefore meltin the rectum or vaginal cavity and release the active compound.

Solid compositions of a similar type may also be employed as fillers insoft and hard-filled gelatin capsules using such excipients as lactoseor milk sugar as well as high molecular weight polyethylene glycols andthe like.

The active compounds can also be in micro-encapsulated form with one ormore excipients as noted above. The solid dosage forms of tablets,dragees, capsules, pills, and granules can be prepared with coatings andshells such as enteric coatings, release controlling coatings and othercoatings well known in the pharmaceutical formulating art. In such soliddosage forms the active compound may be admixed with at least one inertdiluent such as sucrose, lactose or starch. Such dosage forms may alsocomprise, as is normal practice, additional substances other than inertdiluents, e.g., tableting lubricants and other tableting aids such amagnesium stearate and microcrystalline cellulose. In the case ofcapsules, tablets and pills, the dosage forms may also comprisebuffering agents. They may optionally contain opacifying agents and canalso be of a composition that they release the active ingredient(s)only, or preferentially, in a certain part of the intestinal tract,optionally, in a delayed manner. Examples of embedding compositions,which can be used, include polymeric substances and waxes.

Dosage forms for topical or transdermal administration of a compound ofthis invention include ointments, pastes, creams, lotions, gels,powders, solutions, sprays, inhalants or patches. The active componentis admixed under sterile conditions with a pharmaceutically acceptablecarrier and any needed preservatives or buffers as may be required.Ophthalmic formulation, ear drops, eye ointments, powders and solutionsare also contemplated as being within the scope of this invention.

The ointments, pastes, creams and gels may contain, in addition to anactive compound of this invention, excipients such as animal andvegetable fats, oils, waxes, paraffins, starch, tragacanth, cellulosederivatives, polyethylene glycols, silicones, bentonites, silicic acid,talc and zinc oxide, or mixtures thereof.

Powders and sprays can contain, in addition to the compounds of thisinvention, excipients such as lactose, talc, silicic acid, aluminumhydroxide, calcium silicates and polyamide powder, or mixtures of thesesubstances. Sprays can additionally contain customary propellants suchas chlorofluorohydrocarbons.

Transdermal patches have the added advantage of providing controlleddelivery of a compound to the body. Such dosage forms can be made bydissolving or dispensing the compound in the proper medium. Absorptionenhancers can also be used to increase the flux of the compound acrossthe skin. The rate can be controlled by either providing a ratecontrolling membrane or by dispersing the compound in a polymer matrixor gel.

According to the methods of treatment of the present invention, viralinfections are treated or prevented in a subject, such as a human, byadministering to the subject a therapeutically effective amount of acompound of the invention, in such amounts and for such time as isnecessary to achieve the desired result. The term “therapeuticallyeffective amount” of a compound of the invention, as used herein, meansa sufficient amount of the compound so as to decrease the viral load ina subject and/or decrease the subject's HCV symptoms. As is wellunderstood in the medical arts a therapeutically effective amount of acompound of this invention will be at a reasonable benefit/risk ratioapplicable to any medical treatment.

It will be understood, however, that the total daily usage of thecompounds and compositions of the present invention will be decided bythe attending physician within the scope of sound medical judgment. Thespecific therapeutically effective dose level for any particular patientwill depend upon a variety of factors including the disorder beingtreated and the severity of the disorder; the activity of the specificcompound employed; the specific composition employed; the age, bodyweight, general health, sex and diet of the patient; the time ofadministration, route of administration, and rate of excretion of thespecific compound employed; the duration of the treatment; drugs used incombination or contemporaneously with the specific compound employed;and like factors well known in the medical arts.

The total daily dose of the compounds of this invention administered toa human or other animal in single or in divided doses can be in amounts,for example, from 0.01 to 50 mg/kg body weight or more usually from 0.1to 25 mg/kg body weight. Single dose compositions may contain suchamounts or submultiples thereof to make up the daily dose. In general,treatment regimens according to the present invention compriseadministration to a patient in need of such treatment from about 10 mgto about 1000 mg of the compound(s) of this invention per day in singleor multiple doses.

In yet another embodiment, the compounds of the invention may be usedfor the treatment of HCV in humans in monotherapy mode or in acombination therapy (e.g., dual combination, triple combination etc.)mode such as, for example, in combination with antiviral and/orimmunomodulatory agents. Examples of such antiviral and/orimmunomodulatory agents include Ribavirin (from Schering-PloughCorporation, Madison, N.J.) and Levovirin (from ICN Pharmaceuticals,Costa Mesa, Calif.), VP 50406 (from Viropharma, Incorporated, Exton,Pa.), ISIS 14803 (from ISIS Pharmaceuticals, Carlsbad, Calif.),Heptazyme™ (from Ribozyme Pharmaceuticals, Boulder, Colo.), VX 497, andTeleprevir (VX-950) (both from Vertex Pharmaceuticals, Cambridge,Mass.), Thymosin™ (from SciClone Pharmaceuticals, San Mateo, Calif.),Maxamine™ (Maxim Pharmaceuticals, San Diego, Calif.), mycophenolatemofetil (from Hoffman-LaRoche, Nutley, N.J.), interferon (such as, forexample, interferon-alpha, PEG-interferon alpha conjugates) and thelike. “PEG-interferon alpha conjugates” are interferon alpha moleculescovalently attached to a PEG molecule. Illustrative PEG-interferon alphaconjugates include interferon alpha-2a (Roferon™, from Hoffman La-Roche,Nutley, N.J.) in the form of pegylated interferon alpha-2a (e.g., assold under the trade name Pegasys™), interferon alpha-2b (Intron™, fromSchering-Plough Corporation) in the form of pegylated interferonalpha-2b (e.g., as sold under the trade name PEG-Intron™), interferonalpha-2c (BILB 1941, BILN 2061 and Berofor Alpha™, (all from BoehringerIngelheim, Ingelheim, Germany), consensus interferon as defined bydetermination of a consensus sequence of naturally occurring interferonalphas (Infergen™, from Amgen, Thousand Oaks, Calif.). Other suitableanti-HCV agents for use in combination with the present inventioninclude but are not limited to: Yeast-core-NS3 vaccine, EnvelopeVaccine, A-837093 (Abbott Pharmaceuticals), AG0121541 (Pfizer), GS9132(Gilead); HCV-796 (Viropharma), ITMN-191 (Intermune), JTK 003/109 (JapanTobacco Inc.), Lamivudine (EPIVIR) (Glaxo Smith Kline), MK-608 (Merck),R803 (Rigel), ZADAXIN (SciClone Pharmaceuticals); Valopicitabine(Idenix), VGX-410C (Viralgenomix), R1626 (Hoffman La-Roche), andSCH-503034 (Schering Plough Corporation).

Lower or higher doses than those recited above may be required. Specificdosage and treatment regimens for any particular patient will dependupon a variety of factors, including the activity of the specificcompound employed, the age, body weight, general health status, sex,diet, time of administration, rate of excretion, drug combination, theseverity and course of the disease, condition or symptoms, the patient'sdisposition to the disease, condition or symptoms, and the judgment ofthe treating physician.

Unless otherwise defined, all technical and scientific terms used hereinare accorded the meaning commonly known to one with ordinary skill inthe art. All publications, patents, published patent applications, andother references mentioned herein are hereby incorporated by referencein their entirety.

Abbreviations

Abbreviations used in the descriptions of the schemes and the examplesthat follow are:

-   -   aq. for aqueous;    -   CDI for 1,1′-carbonyldiimidizole;    -   DBU for 1,8-diazabicyclo[5.4.0]undec-7-ene;    -   DCM for dichloromethane;    -   DIAD for diisopropyl azodicarboxylate;    -   DIEA for diisopropyl ethylamine;    -   DME for ethylene glycol dimethyl ether;    -   DMF for N,N-dimethyl formamide;    -   ESI for electrospray ionization;    -   Et for ethyl;    -   EtOAc for ethyl acetate;    -   g for gram(s);    -   h for hour(s);    -   HATU for        O-(7-Azabenzotriazole-1-yl)-N,N,N′,N′-tetramethyluronium        hexafluoro-phosphate;    -   HPLC for high-performance liquid chromatography;    -   Me for methyl;    -   MeOH for methanol;    -   mg for milligram(s);    -   min for minute(s);    -   MS for mass spectrometry;    -   NMR for nuclear magnetic resonance;    -   Ph for phenyl;    -   rt for room temperature;    -   THF for tetrahydrofuran;    -   TLC for thin layer chromatography;    -   PPh₃ for triphenylphosphine;    -   tBOC or Boc for tert-butyloxy carbonyl;

Synthetic Methods

The compounds and processes of the present invention will be betterunderstood in connection with the following synthetic schemes thatillustrate the methods by which the compounds of the invention may beprepared.

The targeted analogs were prepared from the common tripeptideintermediates 1-6 and 1-8 and the like. Synthesis toward these versatileintermediates began with the saponification of commercially availableBoc-hydroxyproline methyl ester (1-1) with lithium hydroxide in a 3:1:1mixture of THF/MeOH/water to generate corresponding acid 1-2 (Scheme 1).Subsequent coupling with the cyclopropyl-derived amino acid derivative1-3 exploiting HATU afforded dipeptide 1-4. HCl-mediatedBoc-deprotection in dioxane yielded proline salt 1-5, which was furthercoupled with Boc-tert-L-leucine to give the desired tripeptide 1-6. Itis important to note, that alternative amino acid derivatives can beused in either coupling step to generate tripeptides analogous to 1-6,and therefore ultimately produce multiple alternative pyridazinoneanalogs. Conversely, 1-5 could also be coupled to the olefin-containingamino acid 1-7 delivering a tripeptide that could be cyclized tointermediate 1-8 utilizing the first generation Hoveyda-Grubbs catalyst.

It is important to note, that although only the cis-hydroxyprolineseries is shown in Scheme 1, the trans-hydroxyproline series can becarried through an identical synthetic sequence. Both series are used inorder to generate the targeted HCV inhibitors.

Employing standard Mitsubobu protocols, cis-proline-containingintermediates 1-6 and 1-8 can be transformed to the versatilequinoxaline-containing compounds 2-1 and 2-2. Although this scheme isnot comprehensive, the chemistry portrayed therein serves as a generalguide toward multiple quinoxaline-derived species. For further detailson the Mitsunobu reaction, see O. Mitsunobu, Synthesis 1981, 1-28.

As stated above, both the cis-proline- and trans-proline-derivedintermediates were used in this study. Consequently, epi-1-6 and epi-1-8can both be condensed with CDI, and the resulting product subjected tovarious isoindolines to generate a variety of acyclic and cycliccarbamates, represented by 3-1 and 3-2, respectively. As before, scheme3 is not comprehensive, however the chemistry portrayed therein servesas a general guide toward multiple carbamate-derived species.

Functionalization at the N-terminus can be carried out using a two-stepsequence (Scheme 4) beginning with HCl-mediated Boc-deprotection. Oncethe amine salts (4-1 and 4-2) are generated, they can then be condensedwith an array of electrophiles under basic conditions to producealternative carbamates [—(C═O)—O—R¹], amides [—(C═O)—R¹], ureas[—C(═O)—NH—R²], or sulfonamides [—S(O)₂—R¹, —S(O)₂NHR²], wherein R¹ andR² are as previously defined.

The final steps to the targeted analogs of the present invention includea saponification followed by the formation of the requisite hydrazidemoiety. This strategy was initiated with the treatment of the ethylesters 5-1 and 5-2 with lithium hydroxide in a 3:1:1 mixture ofTHF/MeOH/water (Scheme 5a). Once the carboxylic acids 5-3 and 5-4 wereformed, they could transformed to the desired hydrazide compound in oneof two fashions: (I) the carboxylic acid is condensed with CDI, thensubjected to the fully functionalized hydrazide in the presence of DBU(Scheme 5b), or (II) the carboxylic acid is condensed with CDI,subjected to a 1M solution of hydrazine in THF, then the resulting freeacyl hydrazide is treated with various electrophiles (Scheme 5c).Examples of this methodology are illustrated in, but not limited to, theconversion of compounds represented by structure 5-1 and 5-2 to thehydrazide-derived compounds represented by structures 5-5-5-8.

All references cited herein, whether in print, electronic, computerreadable storage media or other form, are expressly incorporated byreference in their entirety, including but not limited to, abstracts,articles, journals, publications, texts, treatises, internet web sites,databases, patents, and patent publications.

EXAMPLES

The compounds and processes of the present invention will be betterunderstood in connection with the following examples, which are intendedas illustrations only and not to limit the scope of the invention.Various changes and modifications to the disclosed embodiments will beapparent to those skilled in the art and such changes and modificationsincluding, without limitation, those relating to the chemicalstructures, substituents, derivatives, formulations and/or methods ofthe invention may be made without departing from the spirit of theinvention and the scope of the appended claims

(Route 1) Synthesis of the Tri-Peptide Intermediates: (Note: thisSequence was Also Carried Out Using the Trans-Hydroxy Proline CompoundAnalogous to Structure 1a)

Step 1A. To a solution of commercially available cis-L-hydroxyprolinemethyl ester (1a) (1.00 g, 4.1 mmol) in 165 ml of a 3:1:1 mixture ofTHF/MeOH/water at room temperature was added LiOH.H₂O (0.51 g, 12.2mmol). The resulting heterogeneous reaction was stirred at roomtemperature for 14 h, at which time the reaction was concentrated to ˜⅕of its original volume, then acidified with 6 M HCl(aq). This aqueoussolution was then diluted with 20 mL brine and extracted with DCM (4×50mL). The organic washings were combined, washed once with brine, dried(Na₂SO₄), filtered, and concentrated in vacuo. The resulting crudecarboxylic acid 1b was carried on without further purification.

Step 1B. Carboxylic acid 1b (4.08 mmol) was diluted with 50 mL of DCM,cooled to 0° C., then consecutively treated with DIEA (4.1 g, 32.6mmol), cyclopropyl-derived amino-acid hydrochloride salt 1c (0.78 g, 4.1mmol), and HATU (1.9 g, 5.10 mmol). The reaction mixture was allowed towarm to room temperature and closely monitored using mass spectrometricanalysis. Once the reaction was complete, it was transferred to a 250 mLseparatory funnel with 75 mL EtOAc, at which time it was extracted withsaturated aqueous NaHCO₃ (2×20 ml) and brine (2×20 ml). The organicphase was dried over anhydrous Na₂SO₄, filtered, and then concentratedin vacuo. The residue was purified by silica gel flash chromatographyusing gradient elution with hexanes:EtOAc (5:1→3:1→1:1→1:2→1:5) yieldingthe dipeptide 1d (0.826 g, 55%).

MS (ESI) m/z=369.3 (M+H)⁺.

Step 1C. To neat dipeptide 1d was added 20 mL of a 4 M HCl solution indioxane. The resulting mixture was stirred at room temperature for 3 h.Once Boc-deprotection was complete, the excess HCl and organic solventwas removed in vacuo. The resulting amino salt 1e was used without anyfurther purification.

MS (ESI) m/z=269.2 (M+H)⁺.

Step 1D. Amine salt 1e (2.2 mmol) was diluted with 25 mL of DCM, cooledto 0° C., then consecutively treated with DIEA (1.41 g, 11.2 mmol),Boc-tert-L-leucine (0.52 g, 2.2 mmol), and HATU (1.06 g, 2.8 mmol). Thereaction mixture was allowed to warm to room temperature and closelymonitored using mass spectrometric analysis. Once the reaction wascomplete, it was transferred to a 250 mL separatory funnel with 100 mLEtOAc, at which time it was extracted with saturated aqueous NaHCO₃(2×20 ml) and brine (2×20 ml). The organic phase was dried overanhydrous Na₂SO₄, filtered, and then concentrated in vacuo. The residuewas purified by silica gel flash chromatography using gradient elutionwith hexanes:EtOAc (5:1→3:1→1:1→1:2→1:5) yielding the desired tripeptideintermediate 1f (113 g, 93%) as a white solid.

MS (ESI) m/z=482.4 (M+H)⁺.

Step 1E. Amine salt 1e (2.24 mmol) was diluted with 25 mL of DCM, cooledto 0° C., then consecutively treated with DIEA (1.41 g, 11.2 mmol),amino acid 1g (0.61 g, 2.24 mmol), and HATU (1.06 g, 2.80 mmol). Thereaction mixture was allowed to warm to room temperature and closelymonitored using mass spectrometric analysis. Once the reaction wascomplete, it was transferred to a 250 mL separatory funnel with 100 mLEtOAc, at which time it was extracted with saturated aqueous NaHCO₃(2×20 ml) and brine (2×20 ml). The organic phase was dried overanhydrous Na₂SO₄, filtered, and then concentrated in vacuo. The residuewas purified by silica gel flash chromatography using gradient elutionwith hexanes:EtOAc (5:1→3:1→1:1→1:2→1:5) yielding the desired tripeptideintermediate 1h (1.0 g, 97%) as a white solid.

MS (ESI) m/z=544.84 (M+Na)⁺.

Step 1F. A solution of the linear tripeptide 1f (1.5 g, 2.89 mmol) in200 ml dry DCM was deoxygenated by N₂ bubbling (ca. 35 min). Hoveyda's1^(st) generation catalyst (5 mol % eq.) was then added as a solid. Thereaction was refluxed under N₂ atmosphere for 12 h. The solvent wasevaporated and the residue was purified by silica gel flashchromatography using gradient elution with hexanes:EtOAc(9:1→5:1→3:1→1:1→1:2→1:5). The cyclic peptide precursor 1i was isolatedas a white powder (1.2 g, 87%). For further details of the syntheticmethods employed to produce the cyclic peptide precursor 1i, see WO00/059929 (2000).

MS (ESI) m/z=516.28 (M+Na)⁺.

Example 1

Compound of Formula IX, wherein

Step 2A.

To a cooled mixture of macrocyclic precursor 1i,3-(thiophen-2-yl)-1H-quinoxalin-2-one (1.1 equiv.), andtriphenylphosphine (2 equiv.) in THF was added DIAD (2 equiv.) dropwiseat 0° C. The resulting mixture was held at 0° C. for 15 min before beingwarmed to room temperature. After 1 h, the mixture was concentratedunder vacuum and the residue was purified by flash chromatographyeluting with 60% EtOAc in hexanes to give 2a as a clear oil (100 mg,99%).

MS (ESI) m/z=704.4 (M+H)⁺.

H¹-NMR [CDCl₃, δ (ppm)]: 8.6 (d, 1H), 8.0 (d, 1H), 7.8 (d, 1H), 7.6 (m,2H), 7.5 (d, 2H), 7.2 (t, 1H), 7.0 (brs, 1H), 6.0 (brt, 1H), 5.5 (m,1H), 5.3 (brd, 1H), 5.2 (t, 1H), 5.0 (m. 1H), 4.6 (brt, 1H), 4.1-4.3 (m,3H), 3.1 (m, 1H), 5.3 (m, 1H), 2.1-2.3 (m, 2H), 1.3 (brs, 9H), 1.2 (t,3H).

Step 2B.

Compound 2a (82 mg, 0.11 mmol) was treated with HCl (4 M in dioxane, 3mL, 12 mmol). The reaction mixture was stirred at room temperature for 2h until MS showed complete consumption of starting material. The solventwas removed in vacuo and carried directly onto the next step withoutfurther purification.

Step 2C.

The chloroformate reagent 2c was prepared by subjecting 0.22 mmolcyclopentanol in THF (5 ml) to 0.45 mmol of phosgene in toluene (20%).The resulting reaction mixture was stirred at room temperature for 2 hand the solvent was then removed in vacuo. To the residue was added DCMand the resulting mixture was subsequently concentrated in vacuo (repeat2×) yielding chloroformate reagent 2c.

Amine salt 2b (ca. 0.11 mmol) was dissolved in DCM (3 mL) then treatedwith cyclopentyl chloroformate (2c, 0.22 mmol) and DIEA (0.35 mL, 2mmol). The reaction mixture was stirred for 2.5 h. Ethyl acetate (15 mL)was added to the solution, and the resulting reaction mixture wasconsecutively washed with saturated aqueous NaHCO₃ solution, water, andbrine. The organic layer was dried over anhydrous Na₂SO₄. The organicphase was then filtered, concentrated in vacuo and purified by flashchromatography (EtOAc/hexanes 1:2) to give 60.0 mg of the cyclopentylcarbamate 2d.

MS (ESI) m/z 716.31 (M+H)⁺.

Step 2D.

Carbamate 2d (60 mg, from above) was dissolved in THF/MeOH/H₂O (2:1:0.5)and subsequently subjected to lithium hydroxide (10 equiv.) at roomtemperature for 20 h. The excess solvents were evaporated in vacuo, andthe resulting residue was diluted with water and acidified to pH 5. Themixture was extracted with EtOAc (2×). The combined organic extractswere washed once with brine, dried (MgSO₄), filtered and concentrated invacuo to give carboxylic acid 2e, (42.0 mg, 55% for three steps).

MS (ESI) m/z 688.37 (M+H)⁺.

¹³C-NMR (125 MHz, CD₃OD): δ 174.6, 173.5, 173.0, 156.7, 152.9, 141.1,140.0, 139.2, 138.8, 133.4, 130.8, 130.1, 129.3, 128.0, 127.2, 126.7,126.3, 77.5, 76.2, 59.7, 53.3, 52.6, 40.3, 34.8, 34.4, 32.4, 32.2, 32.1,30.8, 27.5, 27.4, 26.4, 23.6, 23.3, 23.0, 22.3.

Step 2E.

In a one-dram vial, carboxylic acid 2e (0.015 g, 0.022 mmol) wasdissolved in 0.75 mL DCM then treated with CDI (5.3 mg, 0.033 mmol). Theresulting mixture was then moved to a 45° C. oil bath and stirred for 1h. After cooling to rt, the vial was opened and nicotinic hydrazide (8.6mg, 0.066 mmol) was added along with DBU (9.2 μL, 0.066 mmol). The vialwas then purged N₂, capped, and moved back to the 45° C. oil bath, whereit was stirred for 3 h. After cooling, the reaction mixture was loadeddirectly onto a plug of SiO₂ and purified via flash chromatography usingEtOAc in hexanes (20%→50%→95%) then 5% MeOH in EtOAc to yield the titlecompound, (10.0 mg, 57%) as a white solid.

MS (ESI) m/z=807.3 (M+H)⁺.

Example 2

Compound of Formula IX, wherein

Step 2E from above was followed using acetic hydrazide instead ofnicotinic hydrazide.

MS (ESI) m/z=744.3 (M+H)⁺.

Example 3

Compound of Formula IX, wherein

Step 2E from above was followed using hydrazine (1.0 M in THF) insteadof nicotinic hydrazide (note: DBU was not needed for this particulartransformation).

MS (ESI) m/z=690.2 (M+H)⁺.

Example 4

Compound of Formula IX, wherein

Step 3A.

Synthesis of the title compound was initiated by the condensation ofalcohol 3a (200 mg, 0.40 mmol—generated using trans-hydroxyproline andthe methodology outlined in Example 1) with CDI (79 mg, 0.49 mmol) in 5mL DCM at rt. Once this coupling was complete as confirmed by MSanalysis, the isoindoline (145 mg, 1.21 mmol) was added and theresulting mixture was stirred overnight. The reaction mixture wasdiluted with DCM (20 mL) and washed with 1N aq. HCl (20 mL) and brine(20 mL). The organic portion was then dried (Na₂SO₄), filtered, andconcentrated in vacuo. The crude oil was purified via flashchromatography (silica gel) using dichloromethane/EtOAc/acetone(60:20:1) as eluent to afford the corresponding carbamate (220 mg, 85%)as a white solid.

Once the carbamate portion was installed, ester hydrolysis was carriedout in standard fashion using LiOH in a THF/MeOH/water (1.5, 0.5, 0.5mL, respectfully) solvent mixture. Upon completion, the reaction mixturewas diluted with 50 mL DCM and 5 mL water, which was acidified with 1Naq. HCl. The layers were separated and the aqueous portion washed threeadditional times with 10 mL DCM. The organic portions were combined andwashed once with brine (20 mL). Finally, the organic layer was dried(Na₂SO₄), filtered, and concentrated in vacuo. The crude acid 3b wascarried on to the coupling step without any further purification.

MS (ESI) m/z=611.3 (M+H)⁺.

Step 3B.

Using 15 mg of carboxylic acid 3b, Step 2E from above was followed usinghydrazine (1.0 M in THF) instead of nicotinic hydrazide (note: DBU wasnot needed for this particular transformation) to give the desiredcompound.

MS (ESI) m/z=643.4 (M+H)⁺.

Example 5

Compound of Formula IX, wherein

Step 4A.

Compound 4 (5 mg, 0.0078 mmol) from above was subjected tocyclopropanecarbonyl chloride (0.8 mg, 0.0078 mmol) in DCM (1 mL) in thepresence of triethylamine (2.4 mg, 0.023 mmol) for 20 min at rt toproduce the desired acylated analog, compound 5.

MS (ESI) m/z=711.2 (M+H)⁺.

Example 6

Compound of Formula IX, wherein

Starting from Compound 3, Step 4a from above was followed usingtert-butylcarbonyl chloride instead of cyclopropanecarbonyl chloride.

MS (ESI) m/z=774.4 (M+H)⁺.

Example 7

Compound of Formula IX, wherein

Starting from Compound 3, Step 4a from above was followed using(S)-tetrahydrofuran-3-chloroformate instead of cyclopropanecarbonylchloride.

MS (ESI) m/z=804.4 (M+H)⁺.

Example 8

Compound of Formula IX, wherein

Starting from Compound 3, Step 4a from above was followed.

MS (ESI) m/z=758.3 (M+H)⁺.

Example 9

Compound of Formula IX, wherein

Starting from Compound 3, Step 4a from above was followed usingmethylchloroformate instead of cyclopropanecarbonyl chloride.

MS (ESI) m/z=748.3 (M+H)⁺.

Example 10

Compound of Formula IX, wherein

Starting from Compound 3, Step 4a from above was followed using phenylisocyanate instead of cyclopropanecarbonyl chloride (note, no exogenousbase was required).

MS (ESI) m/z=809.4 (M+H)⁺.

Example 11

Compound of Formula IX, wherein

Starting from Compound 3, Step 4a from above was followed usingp-nitrobenzoyl chloride instead of cyclopropanecarbonyl chloride.

MS (ESI) m/z=839.4 (M+H)⁺.

Example 114

Compound of Formula XI, wherein

Starting from 1f, Steps 2A, 2D, and 2E from above were followed, usingnaphthalene-2-hydrazide instead of nicotinic hydrazide in Step 2E.

MS (ESI) m/z=832.5 (M+H)⁺.

Example 115

Compound of Formula XI, wherein

Starting from 1f, Steps 2A, 2D, and 2E from above were followed, usingphenyl hydrazide instead of nicotinic hydrazide in Step 2E.

MS (ESI) m/z=782.4 (M+H)⁺.

Example 116

Compound of Formula XI, wherein

Starting from 1f, Steps 2A, 2D, and 2E from above were followed.

MS (ESI) m/z=783.4 (M+H)⁺.

Example 117

Compound of Formula XI, wherein

Starting from 1f, Steps 2A, 2D, and 2E from above were followed, usingfuranyl-2-hydrazide instead of nicotinic hydrazide in Step 2E.

MS (ESI) m/z=772.4 (M+H)⁺.

Example 118

Compound of Formula XI, wherein

Starting from 1f, Steps 2A, 2D, and 2E from above were followed, usingacetic hydrazide instead of nicotinic hydrazide in Step 2E.

MS (ESI) m/z=720.4 (M+H)⁺.

Example 119

Compound of Formula XI, wherein

Starting from 1f, Steps 2A, 2D, and 2E from above were followed, usingthiophenyl-2-hydrazide instead of nicotinic hydrazide in Step 2E.

MS (ESI) m/z=788.4 (M+H)⁺.

Examples 12-113

(Formula IX, Table 3) may be prepared following procedures similar tothose outlined in Examples 1-11.

Wherein A, P*, and V are delineated for each example as set forth inTable 3.

TABLE 3 EXAMPLE # A P* V 12

13

14

15

16

17

18

19

20

21

22

23

24

25

26

27

28

29

30

31

32

33

34

35

36

37

38

39

40

41

42

43

44

45

46

47

48

49

50

51

52

53

54

55

56

57

58

59

60

61

62

63

64

65

66

67

68

69

70

71

72

73

74

75

76

77

78

79

80

81

82

83

84

85

86

87

88

89

90

91

92

93

94

95

96

97

98

99

100

101

102

103

104

105

106

107

108

109

110

111

112

113

Examples 120-226

(Formula X, Table 4) may be prepared following procedures similar tothose outlined in Examples 114-119.

Wherein A, P*, V, and L are delineated for each example as set forth inTable 4:

TABLE 4 EXAMPLE # A P* V L 120

121

122

123

124

125

126

127

128

129

130

131

132

133

134

135

136

137

138

139

140

141

142

143

144

145

146

147

148

149

150

151

152

153

154

155

156

157

158

159

160

161

162

163

164

165

166

167

168

169

170

171

172

173

174

175

176

177

178

179

180

181

182

183

184

185

186

187

188

189

190

191

192

193

194

195

196

197

198

199

200

201

202

203

204

205

206

207

208

209

210

211

212

213

214

215

216

217

218

219

220

221

222

223

224

225

226

The compounds of the present invention exhibit potent inhibitoryproperties against the HCV NS3 protease. The following examples describeassays in which the compounds of the present invention can be tested foranti-HCV effects.

NS3/NS4a Protease Enzyme Assay

HCV protease activity and inhibition is assayed using an internallyquenched fluorogenic substrate. A DABCYL and an EDANS group are attachedto opposite ends of a short peptide. Quenching of the EDANS fluorescenceby the DABCYL group is relieved upon proteolytic cleavage. Fluorescenceis measured with a Molecular Devices Fluoromax (or equivalent) using anexcitation wavelength of 355 nm and an emission wavelength of 485 nm.

The assay is run in Corning white half-area 96-well plates (VWR29444-312 [Corning 3693]) with full-length NS3 HCV protease 1b tetheredwith NS4A cofactor (final enzyme concentration 1 to 15 nM). The assaybuffer is complemented with 10 μM NS4A cofactor Pep 4A (Anaspec 25336 orin-house, MW 1424.8). RET S1(Ac-Asp-Glu-Asp(EDANS)-Glu-Glu-Abu-[COO]Ala-Ser-Lys-(DABCYL)-NH₂,AnaSpec 22991, MW 1548.6) is used as the fluorogenic peptide substrate.The assay buffer contains 50 mM Hepes at pH 7.5, 30 mM NaCl and 10 mMBME. The enzyme reaction is followed over a 30 minutes time course atroom temperature in the absence and presence of inhibitors.

The peptide inhibitors HCV Inh 1 (Anaspec 25345, MW 796.8)Ac-Asp-Glu-Met-Glu-Glu-Cys-OH, [−20° C.] and HCV Inh 2 (Anaspec 25346,MW 913.1) Ac-Asp-Glu-Dif-Cha-Cys-OH, are used as reference compounds.

IC50 values are calculated using XLFit in ActivityBase (IDBS) usingequation 205: y=A+((B−A)/(1+((C/x)̂D))).

Cell-Based Replicon Assay

Quantification of HCV replicon RNA in cell lines (HCV Cell Based Assay).

Cell lines, including Huh-11-7 or Huh 9-13, harboring HCV replicons(Lohmann, et al Science 285:110-113, 1999) are seeded at 5×10³cells/well in 96 well plates and fed media containing DMEM (highglucose), 10% fetal calf serum, penicillin-streptomycin andnon-essential amino acids. Cells are incubated in a 7.5% CO₂ incubatorat 37° C. At the end of the incubation period, total RNA is extractedand purified from cells using Qiagen Rneasy 96 Kit (Catalog No. 74182).To amplify the HCV RNA so that sufficient material can be detected by anHCV specific probe (below), primers specific for HCV (below) mediateboth the reverse transcription of the HCV RNA and the amplification ofthe cDNA by polymerase chain reaction (PCR) using the TaqMan One-StepRT-PCR Master Mix Kit (Applied Biosystems catalog no. 4309169). Thenucleotide sequences of the RT-PCR primers, which are located in theNS5B region of the HCV genome, are the following:

HCV Forward primer “RBNS5bfor” 5′GCTGCGGCCTGTCGAGCT: (SEQ ID NO: 1) HCVReverse primer “RBNS5Brev” 5′CAAGGTCGTCTCCGCATAC. (SEQ ID NO 2)

Detection of the RT-PCR product is accomplished using the AppliedBiosystems (ABI) Prism 7500 Sequence Detection System (SDS) that detectsthe fluorescence that is emitted when the probe, which is labeled with afluorescence reporter dye and a quencher dye, is processed during thePCR reaction. The increase in the amount of fluorescence is measuredduring each cycle of PCR and reflects the increasing amount of RT-PCRproduct. Specifically, quantification is based on the threshold cycle,where the amplification plot crosses a defined fluorescence threshold.Comparison of the threshold cycles of the sample with a known standardprovides a highly sensitive measure of relative template concentrationin different samples (ABI User Bulletin #2 Dec. 11, 1997). The data isanalyzed using the ABI SDS program version 1.7. The relative templateconcentration can be converted to RNA copy numbers by employing astandard curve of HCV RNA standards with known copy number (ABI UserBulletin #2 Dec. 11, 1997).

The RT-PCR product was detected using the following labeled probe:

(SEQ ID NO: 3) 5′ FAM-CGAAGCTCCAGGACTGCACGATGCT-TAMRA FAM = Fluorescencereporter dye. TAMRA = Quencher dye.

The RT reaction is performed at 48° C. for 30 minutes followed by PCR.Thermal cycler parameters used for the PCR reaction on the ABI Prism7500 Sequence Detection System are: one cycle at 95° C., 10 minutesfollowed by 40 cycles each of which include one incubation at 95° C. for15 seconds and a second incubation for 60° C. for 1 minute.

To normalize the data to an internal control molecule within thecellular RNA, RT-PCR is performed on the cellular messenger RNAglyceraldehydes-3-phosphate dehydrogenase (GAPDH). The GAPDH copy numberis very stable in the cell lines used. GAPDH RT-PCR is performed on thesame exact RNA sample from which the HCV copy number is determined. TheGAPDH primers and probes, as well as the standards with which todetermine copy number, are contained in the ABI Pre-Developed TaqManAssay Kit (catalog no. 4310884E). The ratio of HCV/GAPDH RNA is used tocalculate the activity of compounds evaluated for inhibition of HCV RNAreplication.

Activity of Compounds as Inhibitors of HCV Replication (Cell BasedAssay) in Replicon Containing Huh-7 Cell Lines

The effect of a specific anti-viral compound on HCV replicon RNA levelsin Huh-11-7 or 9-13 cells is determined by comparing the amount of HCVRNA normalized to GAPDH (e.g. the ratio of HCV/GAPDH) in the cellsexposed to compound versus cells exposed to the 0% inhibition and the100% inhibition controls. Specifically, cells are seeded at 5×10³cells/well in a 96 well plate and are incubated either with: 1) mediacontaining 1% DMSO (0% inhibition control), 2) 100 international units,IU/ml Interferon-alpha 2b in media/1% DMSO or 3) media/1% DMSOcontaining a fixed concentration of compound. 96 well plates asdescribed above are then incubated at 37° C. for 3 days (primaryscreening assay) or 4 days (IC50 determination). Percent inhibition isdefined as:

% Inhibition=[100−((S−C2)/C1−C2))]×100

-   -   where:    -   S=the ratio of HCV RNA copy number/GAPDH RNA copy number in the        sample;    -   C1=the ratio of HCV RNA copy number/GAPDH RNA copy number in the        0% inhibition control (media/1% DMSO); and    -   C2=the ratio of HCV RNA copy number/GAPDH RNA copy number in the        100% inhibition control (100 IU/ml Interferon-alpha 2b).

The dose-response curve of the inhibitor is generated by adding compoundin serial, three-fold dilutions over three logs to wells starting withthe highest concentration of a specific compound at 10 uM and endingwith the lowest concentration of 0.0 uM. Further dilution series (1 uMto 0.001 uM for example) is performed if the IC50 value is not in thelinear range of the curve. IC50 is determined based on the IDBS ActivityBase program using Microsoft Excel “XL Fit” in which A=100% inhibitionvalue (100 IU/ml Interferon-alpha 2b), B=0% inhibition control value(media/1% DMSO) and C=midpoint of the curve as defined as C=(B−A/2)+A.A, B and C values are expressed as the ratio of HCV RNA/GAPDH RNA asdetermined for each sample in each well of a 96 well plate as describedabove. For each plate the average of 4-6 wells are used to define the100% and 0% inhibition values.

In the above assays, representative compounds are found to haveactivity.

Although the invention has been described with respect to variouspreferred embodiments, it is not intended to be limited thereto, butrather those skilled in the art will recognize that variations andmodifications may be made therein which are within the spirit of theinvention and the scope of the appended claims.

1. A compound represented by Formula I or Formula II, orpharmaceutically acceptable salts, esters, or prodrugs thereof:

Wherein A is selected from the following groups: (i) —(C═O)—O—R¹; (ii)—(C═O)—R¹; (iii) —C(═O)—NH—R²; (iv) —S(O)₂—R¹; (v) —S(O)₂NHR²;  Wherein,R¹ is selected from the following groups: (i) aryl; (ii) substitutedaryl; (iii) heteroaryl; (iv) substituted heteroaryl; (v)heterocycloalkyl; (vi) substituted heterocycloalkyl; (vii) —C₁-C₈ alkyl,optionally containing 0, 1, 2, or 3 heteroatoms selected from O, S, orN; (viii) —C₂-C₈ alkenyl, optionally containing 0, 1, 2, or 3heteroatoms selected from O, S, or N; (ix) —C₂-C₈ alkynyl, optionallycontaining 0, 1, 2, or 3 heteroatoms selected from O, S, or N; (x)substituted —C₁-C₈ alkyl, optionally containing 0, 1, 2, or 3heteroatoms selected from O, S, or N; (xi) substituted —C₂-C₈ alkenyl,optionally containing 0, 1, 2, or 3 heteroatoms selected from O, S, orN; (xii) substituted —C₂-C₈ alkynyl, optionally containing 0, 1, 2, or 3heteroatoms selected from O, S or N; (xiii) —C₃-C₁₂ cycloalkyl; (xiv)substituted —C₃-C₁₂ cycloalkyl; (xv) —C₃-C₁₂ cycloalkenyl; and (xvi)substituted —C₃-C₁₂ cycloalkenyl;  Wherein, R² is selected from thefollowing groups: (i) hydrogen; (ii) —C₁-C₈ alkyl, optionally containing0, 1, 2, or 3 heteroatoms selected from O, S, or N; (iii) —C₂-C₈alkenyl, optionally containing 0, 1, 2, or 3 heteroatoms selected fromO, S, or N; (iv) —C₂-C₈ alkynyl, optionally containing 0, 1, 2, or 3heteroatoms selected from O, S, or N; (v) substituted —C₁-C₈ alkyl,optionally containing 0, 1, 2, or 3 heteroatoms selected from O, S, orN; (vi) substituted —C₂-C₈ alkenyl, optionally containing 0, 1, 2, or 3heteroatoms selected from O, S, or N; (vii) substituted —C₂-C₈ alkynyl,optionally containing 0, 1, 2, or 3 heteroatoms selected from O, S or N;L is selected from the following groups: (i) —C₁-C₈ alkyl, optionallycontaining 0, 1, 2, or 3 heteroatoms selected from O, S, or N; (ii)—C₂-C₈ alkenyl, optionally containing 0, 1, 2, or 3 heteroatoms selectedfrom O, S, or N; (iii) —C₂-C₈ alkynyl, optionally containing 0, 1, 2, or3 heteroatoms selected from O, S or N; (iv) substituted —C₁-C₈ alkyl,optionally containing 0, 1, 2, or 3 heteroatoms selected from O, S, orN; (v) substituted —C₂-C₈ alkenyl, optionally containing 0, 1, 2, or 3heteroatoms selected from O, S, or N; (vi) substituted —C₂-C₈ alkynyl,optionally containing 0, 1, 2, or 3 heteroatoms selected from O, S or N;

=a bond connected to an undefined stereogenic center; X is absent or isselected from the following groups: (i) O; (ii) S; (iii) NR³;  Wherein,R³ is selected from the following groups: (i) hydrogen; (ii) —C₁-C₈alkyl, optionally containing 0, 1, 2, or 3 heteroatoms selected from O,S, or N; (iii) —C₃-C₁₂ cycloalkyl; (iv) substituted —C₃-C₁₂ cycloalkyl;Y is absent or is selected from the following groups: (i) —C₁-C₆ alkyl,optionally containing 0, 1, 2, or 3 heteroatoms selected from O, S, orN, optionally substituted with one or more substituent selected fromhalogen, aryl, substituted aryl, heteroaryl, or substituted heteroaryl;(ii) —C₂-C₆ alkenyl, optionally containing 0, 1, 2, or 3 heteroatomsselected from O, S, or N, optionally substituted with one or moresubstituent selected from halogen, aryl, substituted aryl, heteroaryl,or substituted heteroaryl; (iii) —C₂-C₆ alkynyl, optionally containing0, 1, 2, or 3 heteroatoms selected from O, S, or N, optionallysubstituted with one or more substituent selected from halogen, aryl,substituted aryl, heteroaryl, or substituted heteroaryl; and (iv)—C₃-C₁₂ cycloalkyl, optionally substituted —C₃-C₁₂ cycloalkyl,heterocycloalkyl, substituted heterocycloalkyl; (v) —C═O; (vi) —C═S;(vii) —SO; (viii) —SO₂; Z is selected from the following groups: (i)aryl or alkylaryl optionally substituted with R⁴; (ii) heteroaryl oralkylheteroaryl optionally substituted with R⁴; (iii) —N═CR⁵R⁶; Wherein, R⁴ is selected from the following groups: (i) C₁₋₆ alkyl; (ii)C₃₋₇ cycloalkyl; (iii) C₁₋₆ alkoxy; (iv) C₃₋₇ cycloalkoxy; (v) halo-C₁₋₆alkyl; (vi) CF₃; (vii) mono- or di-halo-C₁₋₆ alkoxy; cyano; (viii) halo;(ix) thioalkyl; (x) hydroxyl; (xi) alkanoyl; (xii) NO₂; (xiii) SH; (xiv)amino; (xv) C₁₋₆ alkylamino; (xvi) di(C₁₋₆) alkylamino; (xvii)di(C₁₋₁₆)alkylamide; (xviii) carboxyl; (xix) (C₁₋₆) carboxyester; (xx)(C₁₋₆) alkylsulfone; (xxi) (C₁₋₆) alkylsulfonamide; (xxii) di(C₁₋₆)alkyl(alkoxy)amine; (xxiii) C₆₋₁₀ aryl; (xxiv) C₇₋₁₄ alkylaryl; or a(xxv) 5-7 membered monocyclic heterocycle;  Wherein, R⁵ and R⁶ areindependently selected from the following groups: (i) aryl; (ii)substituted aryl; (iii) heteroaryl; (iv) substituted heteroaryl; (takentogether with the nitrogen that they are attached to, R⁵ and R⁶ may alsoform a ring) m=0, 1, or 2; n=1, 2, or 3; Q is selected from thefollowing groups: (i) hydrogen; (ii) SR⁷; (iii) —C₁-C₈ alkyl, optionallycontaining 0, 1, 2, or 3 heteroatoms selected from O, S, or N; (iv)—C₂-C₈ alkenyl, optionally containing 0, 1, 2, or 3 heteroatoms selectedfrom O, S, or N; (v) —C₂-C₈ alkynyl, optionally containing 0, 1, 2, or 3heteroatoms selected from O, S or N; (vi) substituted —C₁-C₈ alkyl,optionally containing 0, 1, 2, or 3 heteroatoms selected from O, S, orN; (vii) substituted —C₂-C₈ alkenyl, optionally containing 0, 1, 2, or 3heteroatoms selected from O, S, or N; (viii) substituted —C₂-C₈ alkynylcontaining 0, 1, 2, or 3 heteroatoms selected from O, S or N; (ix)—C₃-C₁₂ cycloalkyl; (x) substituted —C₃-C₁₂ cycloalkyl; (xi) —C₃-C₁₂cycloalkenyl; (xii) substituted —C₃-C₁₂ cycloalkenyl; (xiii)heterocyclic; and (xiv) substituted heterocyclic;  Wherein, R⁷ isselected from the following groups: (i) aryl; (ii) substituted aryl;(iii) heteroaryl; (iv) substituted heteroaryl; (v) heterocycloalkyl;(vi) substituted heterocycloalkyl; (vii) —C₁-C₈ alkyl, optionallycontaining 0, 1, 2, or 3 heteroatoms selected from O, S, or N; (viii)—C₂-C₈ alkenyl, optionally containing 0, 1, 2, or 3 heteroatoms selectedfrom O, S, or N; (ix) —C₂-C₈ alkynyl, optionally containing 0, 1, 2, or3 heteroatoms selected from O, S, or N; (x) substituted —C₁-C₈ alkyl,optionally containing 0, 1, 2, or 3 heteroatoms selected from O, S, orN; (xi) substituted —C₂-C₈ alkenyl, optionally containing 0, 1, 2, or 3heteroatoms selected from O, S, or N; (xii) substituted —C₂-C₈ alkynyl,optionally containing 0, 1, 2, or 3 heteroatoms selected from O, S or N;(xiii) —C₃-C₁₂ cycloalkyl; (xiv) substituted —C₃-C₁₂ cycloalkyl; (xv)—C₃-C₁₂ cycloalkenyl; and (xvi) substituted —C₃-C₁₂ cycloalkenyl; s=0,1, 2, or 3; V and U are independently selected from the followinggroups: (i) hydrogen; (ii) —(C═O)—O—R⁸; (iii) —(C═O)—R⁹; (iv)—C(═O)—NR¹⁰R¹¹; (taken together with the nitrogen that they are attachedto, U and V may also form a ring)  Wherein, R⁸ and R⁹ are selected fromthe following groups: (i) aryl; (ii) substituted aryl; (iii) heteroaryl;(iv) substituted heteroaryl; (v) heterocycloalkyl; (vi) substitutedheterocycloalkyl; (vii) —C₁-C₈ alkyl, optionally containing 0, 1, 2, or3 heteroatoms selected from O, S, or N; (viii) —C₂-C₈ alkenyl,optionally containing 0, 1, 2, or 3 heteroatoms selected from O, S, orN; (ix) —C₂-C₈ alkynyl, optionally containing 0, 1, 2, or 3 heteroatomsselected from O, S, or N; (x) substituted —C₁-C₈ alkyl, optionallycontaining 0, 1, 2, or 3 heteroatoms selected from O, S, or N; (xi)substituted —C₂-C₈ alkenyl, optionally containing 0, 1, 2, or 3heteroatoms selected from O, S, or N; (xii) substituted —C₂-C₈ alkynyl,optionally containing 0, 1, 2, or 3 heteroatoms selected from O, S or N;(xiii) —C₃-C₁₂ cycloalkyl; (xiv) substituted —C₃-C₁₂ cycloalkyl; (xv)—C₃-C₁₂ cycloalkenyl; and (xvi) substituted —C₃-C₁₂ cycloalkenyl; Wherein, R¹⁰ and R¹¹ are selected from the following groups: (i)hydrogen; (ii) aryl; (iii) substituted aryl; (iv) heteroaryl; (v)substituted heteroaryl; (vi) heterocycloalkyl; (vii) substitutedheterocycloalkyl; (viii) —C₁-C₈ alkyl, optionally containing 0, 1, 2, or3 heteroatoms selected from O, S, or N; (ix) —C₂-C₈ alkenyl, optionallycontaining 0, 1, 2, or 3 heteroatoms selected from O, S, or N; (x)—C₂-C₈ alkynyl, optionally containing 0, 1, 2, or 3 heteroatoms selectedfrom O, S, or N; (xi) substituted —C₁-C₈ alkyl, optionally containing 0,1, 2, or 3 heteroatoms selected from O, S, or N; (xii) substituted—C₂-C₈ alkenyl, optionally containing 0, 1, 2, or 3 heteroatoms selectedfrom O, S, or N; (xiii) substituted —C₂-C₈ alkynyl, optionallycontaining 0, 1, 2, or 3 heteroatoms selected from O, S or N; (xiv)—C₃-C₁₂ cycloalkyl; (xv) substituted —C₃-C₁₂ cycloalkyl; (xvi) —C₃-C₁₂cycloalkenyl; and (xvii) substituted —C₃-C₁₂ cycloalkenyl; (takentogether with the nitrogen that they are attached to, R¹⁰ and R¹¹ mayalso form a ring); W is selected from —CH₂—, —O—, —S—, —S(O)₂—, —CO—,—C(O)O—, —C(O)NH—, —CHF—, —CF₂—, aryl, substituted aryl, heteroaryl, andsubstituted heteroaryl;

=either a carbon-carbon single bond or a carbon-carbon double bond j=0,1, 2, 3, or 4; k=1, 2, or
 3. 2. A compound of claim 1, wherein thecompound is of Formula III or Formula IV:

Wherein A, L, j, k, and V are all as previously defined; J is eitherabsent or ═O, S, NR¹², CO, (CO)NR¹², (CO)O, NR¹²(CO), NH(CO)NH, NR¹²SO₂; Wherein, R¹² is selected from the following groups: (i) hydrogen; (ii)—C₁-C₈ alkyl, optionally containing 0, 1, 2, or 3 heteroatoms selectedfrom O, S, or N; (iii) —C₃-C₁₂ cycloalkyl; (iv) substituted —C₃-C₁₂cycloalkyl; R¹³ is selected from aryl, substituted aryl, heteroaryl, andsubstituted heteroaryl; R⁸¹, R⁸², R⁸³, and R⁸⁴ are independently chosenfrom the following groups: (i) hydrogen; (ii) halogen; (iii) —NO₂; (iv)—CN; (v) MR¹⁴, wherein M is absent, or ═O, S, NR¹⁰R¹¹, wherein R¹⁰ andR¹¹ are as previously defined; (vi) aryl; (vii) substituted aryl; (viii)heteroaryl; (ix) substituted heteroaryl; (x) heterocycloalkyl; and (xi)substituted heterocycloalkyl;  Wherein, R¹⁴ is selected from thefollowing groups: (i) hydrogen; (ii) aryl; (iii) substituted aryl; (iv)heteroaryl; (v) substituted heteroaryl; (vi) heterocycloalkyl; (vii)substituted heterocycloalkyl; (viii) —C₁-C₈ alkyl, optionally containing0, 1, 2, or 3 heteroatoms selected from O, S, or N; (ix) —C₂-C₈ alkenyl,optionally containing 0, 1, 2, or 3 heteroatoms selected from O, S, orN; (x) —C₂-C₈ alkynyl, optionally containing 0, 1, 2, or 3 heteroatomsselected from O, S, or N; (xi) substituted —C₁-C₈ alkyl, optionallycontaining 0, 1, 2, or 3 heteroatoms selected from O, S, or N; (xii)substituted —C₂-C₈ alkenyl, optionally containing 0, 1, 2, or 3heteroatoms selected from O, S, or N; (xiii) substituted —C₂-C₈ alkynyl,optionally containing 0, 1, 2, or 3 heteroatoms selected from O, S or N.3. A compound of claim 1, wherein the compound is of Formula V orFormula VIII:

Wherein A, L, j, k, V, R⁸¹, R⁸², R⁸³, and R⁸⁴ are all as previouslydefined.
 4. A compound of claim 1, wherein the compound is of FormulaVII or Formula VIII:

wherein A, L, j, k, V, J, R¹³, R⁸¹, R⁸², R⁸³, and R⁸⁴ are all aspreviously defined;
 5. A compound according to claim 1, which isselected from compounds 1-113 of Formula IX; wherein A, P*(X—Y-Z), and Vare delineated for each example as set forth in Table 1:

TABLE 1 EXAMPLE # A P* V 1

2

3

H 4

H 5

6

7

8

9

10

11

12

13

14

15

16

17

18

19

20

21

22

23

24

25

26

27

28

29

30

31

32

33

34

35

36

37

38

39

40

41

42

43

44

45

46

47

48

49

50

51

52

53

54

55

56

57

58

59

60

61

62

63

64

65

66

67

68

69

70

71

72

73

74

75

76

77

78

79

80

81

82

83

84

85

86

87

88

89

90

91

92

93

94

95

96

97

98

99

100

101

102

103

104

105

106

107

108

109

110

111

112

113


6. A compound according to claim 1, which is selected from compounds114-226 of Formula X; wherein A, P*(X—Y-Z), V, and L are delineated inTable 2:

TABLE 2 EXAMPLE # A P* V L 114

115

116

117

118

119

120

121

122

123

124

125

126

127

128

129

130

131

132

133

134

135

136

137

138

139

140

141

142

143

144

145

146

147

148

149

150

151

152

153

154

155

156

157

158

159

160

161

162

163

164

165

166

167

168

169

170

171

172

173

174

175

176

177

178

179

180

181

182

183

184

185

186

187

188

189

190

191

192

193

194

195

196

197

198

199

200

201

202

203

204

205

206

207

208

209

210

211

212

213

214

215

216

217

218

219

220

221

222

223

224

225

226


7. A pharmaceutical composition comprising therapeutically effectiveamount of a compound according to claims 1, a pharmaceuticallyacceptable salt, ester, or prodrug thereof alone, or in combination witha pharmaceutically acceptable carrier or excipient.
 8. A method oftreating a hepatitis C viral infection in a subject, comprisingadministering to the subject a pharmaceutical composition according toclaim
 6. 9. A method of inhibiting the replication of hepatitis C virus,the method comprising contacting the hepatitis C virus with aninhibitory amount of a compound of claim
 1. 10. The method of claim 7further comprising administering an additional anti-hepatitis C virusagent.
 11. The method of claim 9, wherein said additional anti-hepatitisC virus agent is selected from the group consisting of: α-interferon,β-interferon, ribavarin, and adamantine.
 12. The method of claim 9,wherein said additional anti-hepatitis C virus agent is an inhibitor ofhepatitis C virus helicase, polymerase, metalloprotease, or IRES. 13.Pharmaceutical composition of claim 8 further comprising an additionalanti-hepatitis C virus agent.
 14. A pharmaceutical composition of claim13 wherein said additional anti-hepatitis C virus agent is selected fromthe group consisting of: α-interferon, β-interferon, ribavarin, andadamantine.
 15. Compound of claim 1 wherein said compound is in asubstantially pure form.